token_classification/10-1038_emboj-2009-321.xml

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<article doi="10.1038/emboj.2009.321" abstract="Autophagy, a lysosomal degradation pathway, is essential for homeostasis, development, neurological diseases, and cancer. Regulation of autophagy in human disease is not well understood. Atg9 is a transmembrane protein required for autophagy, and it has been proposed that trafficking of Atg9 may regulate autophagy. Mammalian Atg9 traffics between the TGN and endosomes in basal conditions, and newly formed autophagosomes in response to signals inducing autophagy. We identified p38IP as a new mAtg9 interactor and showed that this interaction is regulated by p38alpha MAPK. p38IP is required for starvation-induced mAtg9 trafficking and autophagosome formation. Manipulation of p38IP and p38alpha alters mAtg9 localization, suggesting p38alpha regulates, through p38IP, the starvation-induced mAtg9 trafficking to forming autophagosomes. Furthermore, we show that p38alpha is a negative regulator of both basal autophagy and starvation-induced autophagy, and suggest that this regulation may be through a direct competition with mAtg9 for binding to p38IP. Our results provide evidence for a link between the MAPK pathway and the control of autophagy through mAtg9 and p38IP."><fig id="1872"><title></title><label>Figure 1</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1872"/><sd-panel panel_id="4836"><b>(A)</b> <span>The <sd-tag id="sdTag504" source="sdapi" category="entity" entity_type="organism" role="assayed" text="AH109" ext_ids="4932" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Saccharomyces cerevisiae" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">AH109</sd-tag> strain was co‐transformed with</span> pGBKT7 <span>containing the C‐terminal (CT) domain of m<sd-tag id="sdTag506" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q674R7///Q7Z3C6" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag> (bait) and</span> pGADT7, or pACT2<span>, containing the CT domain of <sd-tag id="sdTag505" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38IP" ext_ids="Q8NEM7" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38IP</sd-tag> (prey). The interaction was then visualized by</span> <sd-tag id="sdTag7" source="sdapi" category="assay" entity_type="" role="" text="growth" ext_ids="BAO_0002100" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="cell growth assay" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&amp;conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">growth</sd-tag> of co‐transformed yeast on SD‐Trp/Leu/His plates for 3 days at 30°C.<graphic href="https://api.sourcedata.io/file.php?panel_id=4836"/></sd-panel><sd-panel panel_id="4838"><b>(C)</b> <sd-tag id="sdTag466" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HEK293A" ext_ids="CVCL_6910" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293-A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HEK293A</sd-tag> cells were transfected with <sd-tag id="sdTag467" source="sdapi" category="entity" entity_type="protein" role="reporter" text="RFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">RFP</sd-tag><span>-m<sd-tag id="sdTag507" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q674R7///Q7Z3C6" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag> and</span> <sd-tag id="sdTag469" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag470" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38IP" ext_ids="Q8NEM7" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38IP</sd-tag>. <sd-tag id="sdTag471" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag472" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38IP" ext_ids="Q8NEM7" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38IP</sd-tag> was <sd-tag id="sdTag473" source="sdapi" category="assay" entity_type="" role="" text="immunoprecipitated" ext_ids="BAO_0002508" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunoprecipitation" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&amp;conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoprecipitated</sd-tag> with an anti‐<sd-tag id="sdTag474" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag> antibody and the <sd-tag id="sdTag510" source="sdapi" category="assay" entity_type="" role="" text="co‐immunoprecipitated" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">co‐immunoprecipitated</sd-tag> <sd-tag id="sdTag476" source="sdapi" category="entity" entity_type="protein" role="reporter" text="RFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">RFP</sd-tag>-<span>m<sd-tag id="sdTag509" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q674R7///Q7Z3C6" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag></span> was detected with an anti‐<span>m<sd-tag id="sdTag508" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q674R7///Q7Z3C6" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag></span> antibody. <sd-tag id="sdTag479" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag480" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38IP" ext_ids="Q8NEM7" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38IP</sd-tag> was detected with an anti‐<sd-tag id="sdTag481" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag> antibody. Asterisk indicates a non‐specific band.<graphic href="https://api.sourcedata.io/file.php?panel_id=4838"/></sd-panel><sd-panel panel_id="4839">Indirect immunofluorescence analysis of m<sd-tag id="sdTag514" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q7Z3C6///Q674R7" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9A///ATG9B" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag> with (<b>D</b>)<sd-tag id="sdTag16" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-full‐length <sd-tag id="sdTag490" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38IP" ext_ids="55578" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38IP</sd-tag> (<sd-tag id="sdTag21" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-FL), (<b>E</b>)<sd-tag id="sdTag22" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-C‐terminal <sd-tag id="sdTag512" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38IP" ext_ids="55578" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38IP</sd-tag> (<sd-tag id="sdTag24" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-CT), and (<b>F</b>) <sd-tag id="sdTag25" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-N‐terminal <sd-tag id="sdTag511" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38IP" ext_ids="55578" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38IP</sd-tag> (<sd-tag id="sdTag27" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-NT). Bars=5 μm. The co‐localization seen in (D) and (E) was not because of overlap of randomly distributed <sd-tag id="sdTag18" source="sdapi" category="entity" entity_type="subcellular" role="component" text="vesicles" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">vesicles</sd-tag>. Cross‐correlation function (CCF) analysis ( van Steensel <i>et al</i>, 1996) showed a peak and maximum Rp value around Δ<i>X</i>=0, indicating that co‐localization between m<sd-tag id="sdTag513" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q674R7///Q7Z3C6" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag> and <sd-tag id="sdTag29" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38IP" ext_ids="Q8NEM7" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38IP</sd-tag> is positively correlated and non random ( Supplementary Figure S1).<graphic href="https://api.sourcedata.io/file.php?panel_id=4839"/></sd-panel></fig><fig id="1873"><title></title><label>Figure 2</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1873"/><sd-panel panel_id="4840"><b>(A)</b> <sd-tag id="sdTag33" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HEK293A" ext_ids="CVCL_6910" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293-A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HEK293A</sd-tag> cells were transfected with <sd-tag id="sdTag41" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag42" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38IP" ext_ids="Q8NEM7" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38IP</sd-tag> alone or co‐transfected with <sd-tag id="sdTag43" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag44" source="sdapi" category="entity" entity_type="gene" role="component" text="p38IP" ext_ids="55578" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38IP</sd-tag> and <sd-tag id="sdTag34" source="sdapi" category="entity" entity_type="protein" role="reporter" text="RFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">RFP</sd-tag><span>-m<sd-tag id="sdTag515" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q7Z3C6///Q674R7" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9A///ATG9B" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag>, homogenized and subjected to centrifugation, and the resulting</span> post‐nuclear supernatant (PNS) was fractionated by centrifugation at 100 000 <i>g</i> into <sd-tag id="sdTag35" source="sdapi" category="entity" entity_type="subcellular" role="experiment" text="membrane" ext_ids="GO:0016020" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="membrane" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">membrane</sd-tag> pellet and <sd-tag id="sdTag36" source="sdapi" category="entity" entity_type="subcellular" role="experiment" text="cytosol" ext_ids="GO:0005829" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="cytosol" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">cytosol</sd-tag>. Equal protein amounts were resolved by <sd-tag id="sdTag37" source="sdapi" category="assay" entity_type="" role="" text="SDS–PAGE" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SDS-PAGE</sd-tag> and <sd-tag id="sdTag38" source="sdapi" category="assay" entity_type="" role="" text="immunoblotted" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&amp;conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotted</sd-tag> <span>with anti‐<sd-tag id="sdTag516" source="sdapi" category="entity" entity_type="protein" role="assayed" text="mannose‐6‐phosphate receptor" ext_ids="P20645" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="M6PR" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">mannose‐6‐phosphate receptor</sd-tag> (</span><sd-tag id="sdTag39" source="sdapi" category="entity" entity_type="protein" role="assayed" text="MPR" ext_ids="P20645" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="M6PR" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">MPR</sd-tag><span>) antibody as a control for membrane‐associated proteins, anti‐<sd-tag id="sdTag518" source="sdapi" category="entity" entity_type="protein" role="assayed" text="superoxide dismutase" ext_ids="P08294///P04179///P00441" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="SOD3///SOD2///SOD1" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">superoxide dismutase</sd-tag> (</span><sd-tag id="sdTag40" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SOD" ext_ids="P08294///P04179///P00441" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="SOD3///SOD2///SOD1" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">SOD</sd-tag><span>) antibody as a control for cytosolic proteins, anti‐m<sd-tag id="sdTag519" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Atg9" ext_ids="285973///79065" norm_text="" ext_dbs="NCBI gene///NCBI gene" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene/">Atg9</sd-tag> antibody, and anti‐</span><sd-tag id="sdTag48" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag> antibody. <sd-tag id="sdTag49" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag50" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38IP" ext_ids="Q8NEM7" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38IP</sd-tag> levels were quantified using ImageJ software and plotted relative to <sd-tag id="sdTag51" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag52" source="sdapi" category="entity" entity_type="gene" role="component" text="p38IP" ext_ids="55578" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38IP</sd-tag> in the PNS (<i>n</i>=4) (<sup>**</sup><i>P</i>=0.0051).<graphic href="https://api.sourcedata.io/file.php?panel_id=4840"/></sd-panel><sd-panel panel_id="4841"><b>(B)</b> <sd-tag id="sdTag53" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HEK293" ext_ids="CVCL_0045" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HEK293</sd-tag> <span>cells were depleted of m<sd-tag id="sdTag521" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Atg9" ext_ids="285973///79065" norm_text="" ext_dbs="NCBI gene///NCBI gene" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene/">Atg9</sd-tag> using siRNA, then transfected with</span> <sd-tag id="sdTag58" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag59" source="sdapi" category="entity" entity_type="gene" role="component" text="p38IP" ext_ids="55578" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38IP</sd-tag> and incubated in full medium or <sd-tag id="sdTag54" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> (data not shown). The asterisk indicates <span>m<sd-tag id="sdTag522" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q674R7///Q7Z3C6" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag></span><span>‐depleted cells. Bar is 5 μM. The intensity of the nuclear versus cytosolic fluorescence in control and cells depleted of m<sd-tag id="sdTag523" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q7Z3C6///Q674R7" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9A///ATG9B" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag> under fed and starvation conditions was analysed using ImageJ software (</span><sup>***</sup><i>P</i>=0.0001, Students <i>t</i>‐test)<graphic href="https://api.sourcedata.io/file.php?panel_id=4841"/></sd-panel><sd-panel panel_id="4842"><b>(C)</b> <sd-tag id="sdTag68" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HEK293A" ext_ids="CVCL_6910" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293-A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HEK293A</sd-tag> cells were transfected with <sd-tag id="sdTag69" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag70" source="sdapi" category="entity" entity_type="gene" role="component" text="p38IP" ext_ids="55578" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38IP</sd-tag> and processed for indirect immunofluorescence. <sd-tag id="sdTag71" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag72" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38IP" ext_ids="Q8NEM7" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38IP</sd-tag> was detected with an anti‐<sd-tag id="sdTag73" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag> <span>antibody and endogenous m<sd-tag id="sdTag524" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q674R7///Q7Z3C6" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag> was detected with an anti‐m<sd-tag id="sdTag525" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q674R7///Q7Z3C6" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag> antibody.</span> <sd-tag id="sdTag76" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag77" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38IP" ext_ids="Q8NEM7" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38IP</sd-tag> localizes to the <sd-tag id="sdTag66" source="sdapi" category="entity" entity_type="subcellular" role="component" text="nucleus" ext_ids="GO:0005634" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="nucleus" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">nucleus</sd-tag> <span>and <sd-tag id="sdTag528" source="sdapi" category="entity" entity_type="subcellular" role="component" text="cytoplasmic" ext_ids="GO:0005737" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="cytoplasm" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">cytoplasmic</sd-tag> puncta in full medium or</span> <sd-tag id="sdTag78" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag>. <sd-tag id="sdTag79" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag80" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38IP" ext_ids="Q8NEM7" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38IP</sd-tag> <span>co‐localizes with m<sd-tag id="sdTag526" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q674R7///Q7Z3C6" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag> in the periphery of the cell (arrowheads). Bars (upper panels) 5 μM, (lower panels) 1 μM.</span><graphic href="https://api.sourcedata.io/file.php?panel_id=4842"/></sd-panel><sd-panel panel_id="4843"><b>(D)</b> Anti‐<span>m<sd-tag id="sdTag529" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Atg9" ext_ids="285973///79065" norm_text="" ext_dbs="NCBI gene///NCBI gene" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene/">Atg9</sd-tag></span> antibody was used to <sd-tag id="sdTag83" source="sdapi" category="assay" entity_type="" role="" text="immunoprecipitate" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">immunoprecipitate</sd-tag> <span>m<sd-tag id="sdTag531" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q674R7///Q7Z3C6" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag></span> from <sd-tag id="sdTag87" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HEK293" ext_ids="CVCL_0045" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HEK293</sd-tag> cells transfected with <sd-tag id="sdTag88" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag496" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38IP" ext_ids="Q8NEM7" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38IP</sd-tag> alone or with <sd-tag id="sdTag90" source="sdapi" category="entity" entity_type="protein" role="reporter" text="RFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">RFP</sd-tag>-<span>m<sd-tag id="sdTag530" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q674R7///Q7Z3C6" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag></span> and <sd-tag id="sdTag92" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag93" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38IP" ext_ids="Q8NEM7" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38IP</sd-tag>. A total of 5% of lysates or immunoprecipitates were probed with anti‐<sd-tag id="sdTag94" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag> or anti‐<span>m<sd-tag id="sdTag532" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q674R7///Q7Z3C6" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag></span> antibodies. The left and right hand bottom panels were obtained from the same <sd-tag id="sdTag84" source="sdapi" category="assay" entity_type="" role="" text="immunoblot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&amp;conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblot</sd-tag>, but exposed for different times to allow the endogenous <span>m<sd-tag id="sdTag533" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q674R7///Q7Z3C6" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag></span> to be visualized better.<graphic href="https://api.sourcedata.io/file.php?panel_id=4843"/></sd-panel><sd-panel panel_id="4844"><b>(E)</b> <sd-tag id="sdTag102" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HEK293" ext_ids="CVCL_0045" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HEK293</sd-tag> cells were transfected with control or <sd-tag id="sdTag103" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38IP" ext_ids="55578" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38IP</sd-tag> siRNA. At <sd-pretag id="sdPretag14961398132080" parent-tag-id="590">72 h</sd-pretag> after transfection, cells were incubated in either full medium or <sd-tag id="sdTag104" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> for 2 h, then fixed and immunostained <span>to detect endogenous m<sd-tag id="sdTag534" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q7Z3C6///Q674R7" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9A///ATG9B" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag> localization. m<sd-tag id="sdTag535" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q7Z3C6///Q674R7" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9A///ATG9B" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag> localization was quantified by visually scoring cells for either a juxta‐nuclear or dispersed phenotype. Bars=5 μm (data are represented as mean±s.e.m. of 200 cells,</span> <sup>*</sup><i>P</i>=0.0413, Student's <i>t</i>‐test).<graphic href="https://api.sourcedata.io/file.php?panel_id=4844"/></sd-panel></fig><fig id="1874"><title></title><label>Figure 3</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1874"/><sd-panel panel_id="4845"><b>(A)</b> <sd-tag id="sdTag108" source="sdapi" category="entity" entity_type="cell_line" role="component" text="293" ext_ids="CVCL_0045" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">293</sd-tag>/<sd-tag id="sdTag109" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag110" source="sdapi" category="entity" entity_type="gene" role="component" text="LC3" ext_ids="440738///81631///84557" norm_text="" ext_dbs="NCBI gene///NCBI gene///NCBI gene" in_caption="True" ext_names="MAP1LC3C///MAP1LC3B///MAP1LC3A" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene/">LC3</sd-tag> cells were transfected with control or <sd-tag id="sdTag113" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38IP" ext_ids="55578" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38IP</sd-tag> siRNA. At <sd-pretag id="sdPretag14961398246130" parent-tag-id="590">72 h</sd-pretag> after transfection, cells were incubated in either full medium (FM), full medium with <sd-tag id="sdTag111" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="leupeptin" ext_ids="CHEBI:6426" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Leupeptin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">leupeptin</sd-tag> (FM Leu), <sd-tag id="sdTag115" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> (ES), or <sd-tag id="sdTag116" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> with <sd-tag id="sdTag117" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="leupeptin" ext_ids="CHEBI:6426" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Leupeptin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">leupeptin</sd-tag> (ES Leu) for 2 h. FM Leu (data not shown) is identical to FM alone. Quantification of <sd-tag id="sdTag119" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag120" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9BXW4///Q9H492///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3C///MAP1LC3A///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag>‐positive <sd-tag id="sdTag112" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="autophagosome" ext_ids="GO:0005776" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="autophagic vacuole" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">autophagosome</sd-tag> number was performed by counting in a blinded experiment. Bars=5 μm (data are represented as mean±s.e.m. of 60 cells, <sup>***</sup><i>P</i>=0.0002, Students <i>t</i>‐test).<graphic href="https://api.sourcedata.io/file.php?panel_id=4845"/></sd-panel><sd-panel panel_id="4846"><b>(B)</b> <sd-tag id="sdTag123" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HEK293A" ext_ids="CVCL_6910" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293-A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HEK293A</sd-tag> cells were transfected with siRNA control, siRNA for <sd-tag id="sdTag124" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38IP" ext_ids="55578" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38IP</sd-tag>, and siRNA for <sd-tag id="sdTag122" source="sdapi" category="entity" entity_type="gene" role="intervention" text="ULK1" ext_ids="8408" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ULK1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">ULK1</sd-tag>. Cell lysates were analysed by <sd-tag id="sdTag125" source="sdapi" category="assay" entity_type="" role="" text="SDS–PAGE" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SDS-PAGE</sd-tag> for endogenous <sd-tag id="sdTag126" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9BXW4///Q9H492///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3C///MAP1LC3A///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag> lipidation using an anti‐<sd-tag id="sdTag130" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9BXW4///Q9H492///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3C///MAP1LC3A///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag> antibody, or <sd-tag id="sdTag128" source="sdapi" category="assay" entity_type="" role="" text="immunoblotted" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&amp;conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotted</sd-tag> with anti‐<sd-tag id="sdTag129" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ULK1" ext_ids="O75385" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ULK1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ULK1</sd-tag> antibody.<graphic href="https://api.sourcedata.io/file.php?panel_id=4846"/></sd-panel><sd-panel panel_id="4847"><b>(C)</b> <sd-tag id="sdTag133" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HeLa" ext_ids="CVCL_0030" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HeLa" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HeLa</sd-tag> cells were transfected with siRNA control, and siRNA for <sd-tag id="sdTag136" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38IP" ext_ids="55578" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38IP</sd-tag>. After incubation for 2 h as in (A), samples were <sd-tag id="sdTag138" source="sdapi" category="assay" entity_type="" role="" text="immunoblotted" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&amp;conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotted</sd-tag> with anti‐<sd-tag id="sdTag139" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9BXW4///Q9H492///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3C///MAP1LC3A///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag> and anti‐<sd-tag id="sdTag134" source="sdapi" category="entity" entity_type="protein" role="normalizing" text="actin" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">actin</sd-tag> <span>antibodies. Endogenous <sd-tag id="sdTag538" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9BXW4///Q9H492///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3C///MAP1LC3A///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag>II/<sd-tag id="sdTag539" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9BXW4///Q9H492///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3C///MAP1LC3A///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag>I levels were quantified and the ratio presented as arbitrary units. In (B),</span> <i>n</i>=4, <sup>***</sup><i>P</i>=0.0001 and <sup>*</sup><i>P</i>=0.0324, Students <i>t</i>‐test. In (C), data are representative of two experiments.<graphic href="https://api.sourcedata.io/file.php?panel_id=4847"/></sd-panel><sd-panel panel_id="4848"><b>(D)</b> <sd-tag id="sdTag148" source="sdapi" category="entity" entity_type="cell_line" role="component" text="293" ext_ids="CVCL_0045" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">293</sd-tag>/<sd-tag id="sdTag149" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag150" source="sdapi" category="entity" entity_type="gene" role="component" text="LC3" ext_ids="440738///81631///84557" norm_text="" ext_dbs="NCBI gene///NCBI gene///NCBI gene" in_caption="True" ext_names="MAP1LC3C///MAP1LC3B///MAP1LC3A" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene/">LC3</sd-tag> cells were transfected with siRNA control or siRNA <sd-tag id="sdTag151" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38IP" ext_ids="55578" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38IP</sd-tag>, and incubated in full medium, or for 2 and 4 h in <sd-tag id="sdTag152" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag>. Samples were <sd-tag id="sdTag153" source="sdapi" category="assay" entity_type="" role="" text="immunoblotted" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&amp;conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotted</sd-tag> with anti‐<sd-tag id="sdTag145" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p62" ext_ids="Q13501" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SQSTM1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p62</sd-tag> antibodies, and <sd-tag id="sdTag154" source="sdapi" category="entity" entity_type="protein" role="normalizing" text="actin" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">actin</sd-tag> as a loading control.<graphic href="https://api.sourcedata.io/file.php?panel_id=4848"/></sd-panel><sd-panel panel_id="4849"><b>(E)</b> <sd-tag id="sdTag159" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HEK293A" ext_ids="CVCL_6910" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293-A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HEK293A</sd-tag> cells were transfected with <sd-tag id="sdTag160" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag161" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38IP" ext_ids="55578" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38IP</sd-tag> or <sd-tag id="sdTag155" source="sdapi" category="entity" entity_type="protein" role="reporter" text="myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">myc</sd-tag>-<sd-tag id="sdTag162" source="sdapi" category="entity" entity_type="gene" role="intervention" text="ULK1" ext_ids="8408" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ULK1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">ULK1</sd-tag> C‐terminal domain (CTD). After 24 h, cells were labelled with <sd-tag id="sdTag157" source="sdapi" category="entity" entity_type="molecule" role="reporter" text="[14C]valine" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">[<sup>14</sup>C]valine</sd-tag> as described in Material and Methods section, and incubated in either full medium or <sd-tag id="sdTag163" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> for 2 h. Cells were then collected and analysed for <sd-tag id="sdTag158" source="sdapi" category="assay" entity_type="" role="" text="long‐lived protein degradation" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">long‐lived protein degradation</sd-tag> (data are represented as mean±s.e.m. of triplicates, representative of two experiments, <sd-tag id="sdTag165" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> mock versus <sd-tag id="sdTag166" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> <sd-tag id="sdTag167" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag168" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38IP" ext_ids="55578" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38IP</sd-tag> (<sup>***</sup><i>P</i>=0.036); <sd-tag id="sdTag169" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> mock versus <sd-tag id="sdTag170" source="sdapi" category="entity" entity_type="gene" role="intervention" text="ULK1" ext_ids="8408" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ULK1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">ULK1</sd-tag> CTD (<sup>***</sup><i>P</i>=0.0001); Students <i>t</i>‐test).<graphic href="https://api.sourcedata.io/file.php?panel_id=4849"/></sd-panel></fig><fig id="1875"><title></title><label>Figure 4</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1875"/><sd-panel panel_id="4850"><b>(A)</b> <sd-tag id="sdTag176" source="sdapi" category="entity" entity_type="cell_line" role="component" text="293" ext_ids="CVCL_0045" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">293</sd-tag>/<sd-tag id="sdTag177" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag178" source="sdapi" category="entity" entity_type="gene" role="component" text="LC3" ext_ids="440738///81631///84557" norm_text="" ext_dbs="NCBI gene///NCBI gene///NCBI gene" in_caption="True" ext_names="MAP1LC3C///MAP1LC3B///MAP1LC3A" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene/">LC3</sd-tag> cells were treated with 10 μM <sd-tag id="sdTag171" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="anisomycin" ext_ids="CHEBI:40748" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="(-)-anisomycin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">anisomycin</sd-tag> for 30 min, or exposed to <span><sd-tag id="sdTag544" source="sdapi" category="physical" entity_type="" role="" text="UV irradiation" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">UV irradiation</sd-tag></span> for 3 min followed by a 40‐min recovery and incubation in full medium, full medium with <sd-tag id="sdTag179" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="leupeptin" ext_ids="CHEBI:6426" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Leupeptin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">leupeptin</sd-tag>, <sd-tag id="sdTag180" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag>, or <sd-tag id="sdTag181" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> with <sd-tag id="sdTag182" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="leupeptin" ext_ids="CHEBI:6426" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Leupeptin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">leupeptin</sd-tag> for 2 h. Cells were then fixed and visualized by <sd-tag id="sdTag175" source="sdapi" category="assay" entity_type="" role="" text="confocal microscopy" ext_ids="BAO_0000453" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="confocal microscopy" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&amp;conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">confocal microscopy</sd-tag>. <sd-tag id="sdTag184" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag185" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9BXW4///Q9H492///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3C///MAP1LC3A///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag>‐positive structures per cell were quantified as in Figure 3A. Bars=5 μm. Data are represented as mean±s.e.m. <i>n</i>=60 cells, mock versus <sd-tag id="sdTag186" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="anisomycin" ext_ids="CHEBI:40748" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="(-)-anisomycin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">anisomycin</sd-tag> <sd-tag id="sdTag187" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> (<sup>***</sup><i>P</i>=0.0001); mock versus UV <sd-tag id="sdTag188" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> (<sup>***</sup><i>P</i>=0.0001); mock versus <sd-tag id="sdTag189" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="anisomycin" ext_ids="CHEBI:40748" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="(-)-anisomycin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">anisomycin</sd-tag> <sd-tag id="sdTag190" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> with <sd-tag id="sdTag191" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="leupeptin" ext_ids="CHEBI:6426" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Leupeptin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">leupeptin</sd-tag> (<sup>***</sup><i>P</i>=0.0001); mock versus <sd-tag id="sdTag192" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> with <sd-tag id="sdTag193" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="leupeptin" ext_ids="CHEBI:6426" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Leupeptin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">leupeptin</sd-tag> UV (<sup>***</sup><i>P</i>=0.0001). All analysed using Student's <i>t</i>-test.<graphic href="https://api.sourcedata.io/file.php?panel_id=4850"/></sd-panel><sd-panel panel_id="4851"><b>(B)</b> <sd-tag id="sdTag196" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HEK293A" ext_ids="CVCL_6910" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293-A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HEK293A</sd-tag> cells were pretreated with 10 μM <sd-tag id="sdTag197" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="anisomycin" ext_ids="CHEBI:40748" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="(-)-anisomycin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">anisomycin</sd-tag> for 30 min<span>, or <sd-tag id="sdTag546" source="sdapi" category="physical" entity_type="" role="" text="UV irradiation" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">UV irradiation</sd-tag> for</span> 3 min followed by a 40‐min recovery, before incubation in <sd-tag id="sdTag202" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> for 2 h. Images were analysed as in Figure 2E. Bars=5 μm. Data are represented as mean±s.e.m. of 60 cells, <sd-tag id="sdTag204" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> mock versus <sd-tag id="sdTag205" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> <sd-tag id="sdTag206" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="anisomycin" ext_ids="CHEBI:40748" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="(-)-anisomycin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">anisomycin</sd-tag> (<sup>*</sup><i>P</i>=0.0142); <sd-tag id="sdTag207" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> mock versus <sd-tag id="sdTag208" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> UV (<sup>*</sup><i>P</i>=0.0150); Student's <i>t</i>‐test.<graphic href="https://api.sourcedata.io/file.php?panel_id=4851"/></sd-panel></fig><fig id="1876"><title></title><label>Figure 5</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1876"/><sd-panel panel_id="4852"><b>(A)</b> <sd-tag id="sdTag214" source="sdapi" category="entity" entity_type="cell_line" role="component" text="293" ext_ids="CVCL_0045" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">293</sd-tag>/<sd-tag id="sdTag215" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag216" source="sdapi" category="entity" entity_type="gene" role="component" text="LC3" ext_ids="440738///81631///84557" norm_text="" ext_dbs="NCBI gene///NCBI gene///NCBI gene" in_caption="True" ext_names="MAP1LC3C///MAP1LC3B///MAP1LC3A" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene/">LC3</sd-tag> cells were pre‐treated with 10 μM <sd-tag id="sdTag217" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="anisomycin" ext_ids="CHEBI:40748" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="(-)-anisomycin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">anisomycin</sd-tag> for 30 min before incubation in either full medium or <sd-tag id="sdTag219" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag>. Cell lysates were analysed by <sd-tag id="sdTag220" source="sdapi" category="assay" entity_type="" role="" text="SDS–PAGE" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SDS-PAGE</sd-tag> for <sd-tag id="sdTag221" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag222" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9BXW4///Q9H492///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3C///MAP1LC3A///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag> lipidation using an anti‐<sd-tag id="sdTag223" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9BXW4///Q9H492///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3C///MAP1LC3A///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag> antibody. The membrane was also probed with anti‐<sd-tag id="sdTag212" source="sdapi" category="entity" entity_type="protein" role="normalizing" text="β tubulin" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">β tubulin</sd-tag>. The <sd-tag id="sdTag225" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag226" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9BXW4///Q9H492///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3C///MAP1LC3A///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag> lipidation was quantified as the amount of <sd-tag id="sdTag227" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-LC3II/<sd-tag id="sdTag229" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-LC3I (data are represented as mean±s.e.m. of triplicates, <i>P</i>=0.0091, Student's <i>t</i>‐test).<graphic href="https://api.sourcedata.io/file.php?panel_id=4852"/></sd-panel><sd-panel panel_id="4853"><b>(B)</b> <sd-tag id="sdTag232" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HEK293A" ext_ids="CVCL_6910" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293-A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HEK293A</sd-tag> cells labelled with <sd-tag id="sdTag231" source="sdapi" category="entity" entity_type="molecule" role="reporter" text="[14C]valine" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">[<sup>14</sup>C]valine</sd-tag>, as described in Material and Methods section, were pretreated with 10 μM <sd-tag id="sdTag233" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="anisomycin" ext_ids="CHEBI:40748" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="(-)-anisomycin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">anisomycin</sd-tag> for 30 min before incubation in either full medium, <sd-tag id="sdTag235" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag>, or <sd-tag id="sdTag236" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> with <sd-tag id="sdTag237" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="leupeptin" ext_ids="CHEBI:6426" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Leupeptin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">leupeptin</sd-tag> for 2 h. Cells were then analysed for <sd-tag id="sdTag239" source="sdapi" category="assay" entity_type="" role="" text="long‐lived protein degradation" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">long‐lived protein degradation</sd-tag> (data are represented as mean±s.e.m. of triplicates, representative of two experiments, <sd-tag id="sdTag240" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> untreated versus <sd-tag id="sdTag241" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="anisomycin" ext_ids="CHEBI:40748" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="(-)-anisomycin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">anisomycin</sd-tag> treated (<i>P</i>=0.0363); <sd-tag id="sdTag242" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> untreated versus <sd-tag id="sdTag243" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="leupeptin" ext_ids="CHEBI:6426" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Leupeptin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">leupeptin</sd-tag> treated (<i>P</i>=0.0001), Student's <i>t</i>‐test).<graphic href="https://api.sourcedata.io/file.php?panel_id=4853"/></sd-panel><sd-panel panel_id="4854"><b>(C)</b> <sd-tag id="sdTag246" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HEK293" ext_ids="CVCL_0045" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HEK293</sd-tag> cells were transfected with control siRNA or siRNA for <sd-tag id="sdTag247" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38IP" ext_ids="55578" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38IP</sd-tag>. After 24‐h incubation, control or <sd-tag id="sdTag248" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38IP" ext_ids="55578" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38IP</sd-tag>‐depleted cells were pre‐treated with <sd-tag id="sdTag249" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="anisomycin" ext_ids="CHEBI:40748" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="(-)-anisomycin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">anisomycin</sd-tag>, followed by a 2‐h incubation with full medium, full medium with <sd-tag id="sdTag250" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="leupeptin" ext_ids="CHEBI:6426" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Leupeptin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">leupeptin</sd-tag>, <sd-tag id="sdTag251" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag>, or <sd-tag id="sdTag252" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> with <sd-tag id="sdTag253" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="leupeptin" ext_ids="CHEBI:6426" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Leupeptin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">leupeptin</sd-tag>. Cell lysates were analysed with anti‐<sd-tag id="sdTag254" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38α" ext_ids="Q16539" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="MAPK14" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38α</sd-tag> or anti‐<sd-tag id="sdTag255" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9BXW4///Q9H492///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3C///MAP1LC3A///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag> <span>antibodies. <sd-tag id="sdTag551" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9BXW4///Q9H492///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3C///MAP1LC3A///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag>II/<sd-tag id="sdTag552" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9BXW4///Q9H492///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3C///MAP1LC3A///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag>I levels were quantified and presented normalized to control, untreated cells incubated in</span> <sd-tag id="sdTag258" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag>. Data are representative of two experiments.<graphic href="https://api.sourcedata.io/file.php?panel_id=4854"/></sd-panel><sd-panel panel_id="4855"><b>(D)</b> <sd-tag id="sdTag261" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HEK293A" ext_ids="CVCL_6910" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293-A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HEK293A</sd-tag> cells transiently transfected with <sd-tag id="sdTag262" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>‐<sd-tag id="sdTag263" source="sdapi" category="entity" entity_type="gene" role="component" text="p38IP" ext_ids="55578" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38IP</sd-tag> and <sd-tag id="sdTag264" source="sdapi" category="entity" entity_type="protein" role="reporter" text="RFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">RFP</sd-tag><span>‐m<sd-tag id="sdTag554" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Atg9" ext_ids="285973///79065" norm_text="" ext_dbs="NCBI gene///NCBI gene" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene/">Atg9</sd-tag> were treated with</span> <sd-tag id="sdTag266" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="anisomycin" ext_ids="CHEBI:40748" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="(-)-anisomycin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">anisomycin</sd-tag> for 30 min <span>and m<sd-tag id="sdTag555" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q7Z3C6///Q674R7" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9A///ATG9B" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag> was</span> <sd-tag id="sdTag260" source="sdapi" category="assay" entity_type="" role="" text="immunoprecipitated" ext_ids="BAO_0002508" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunoprecipitation" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&amp;conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoprecipitated</sd-tag>. 5% of the lysates or the immunoprecipitates were probed with anti‐<sd-tag id="sdTag269" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag> and anti‐mAtg9 antibodies (data is representative of 3 experiments, <sup>*</sup><i>P</i>=0.0332).<graphic href="https://api.sourcedata.io/file.php?panel_id=4855"/></sd-panel></fig><fig id="1877"><title></title><label>Figure 6</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1877"/><sd-panel panel_id="4856"><b>(A)</b> <sd-tag id="sdTag276" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HEK293A" ext_ids="CVCL_6910" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293-A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HEK293A</sd-tag> cells were transfected with <sd-tag id="sdTag277" source="sdapi" category="entity" entity_type="protein" role="reporter" text="RFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">RFP</sd-tag><span>-m<sd-tag id="sdTag556" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Atg9" ext_ids="285973///79065" norm_text="" ext_dbs="NCBI gene///NCBI gene" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene/">Atg9</sd-tag> and</span> <sd-tag id="sdTag273" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag>-<sd-tag id="sdTag274" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38" ext_ids="1432///5603///6300///5600" norm_text="" ext_dbs="NCBI gene///NCBI gene///NCBI gene///NCBI gene" in_caption="True" ext_names="MAPK14///MAPK13///MAPK12///MAPK11" ext_tax_ids="9606///9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene/">p38</sd-tag> (lanes 2 and 7), <sd-tag id="sdTag279" source="sdapi" category="entity" entity_type="protein" role="reporter" text="RFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">RFP</sd-tag><span>-m<sd-tag id="sdTag557" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q7Z3C6///Q674R7" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9A///ATG9B" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag>,</span> <sd-tag id="sdTag281" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag>-<sd-tag id="sdTag282" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38" ext_ids="1432///5603///6300///5600" norm_text="" ext_dbs="NCBI gene///NCBI gene///NCBI gene///NCBI gene" in_caption="True" ext_names="MAPK14///MAPK13///MAPK12///MAPK11" ext_tax_ids="9606///9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene/">p38</sd-tag> and <sd-tag id="sdTag283" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag284" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38IP" ext_ids="55578" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38IP</sd-tag> (lanes 3 and 8), or <sd-tag id="sdTag285" source="sdapi" category="entity" entity_type="protein" role="reporter" text="RFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">RFP</sd-tag><span>-m<sd-tag id="sdTag558" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q674R7///Q7Z3C6" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag> and</span> <sd-tag id="sdTag287" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag288" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38IP" ext_ids="55578" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38IP</sd-tag> (lanes 4 and 9). <sd-tag id="sdTag289" source="sdapi" category="entity" entity_type="protein" role="reporter" text="RFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">RFP</sd-tag><span>-m<sd-tag id="sdTag563" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q674R7///Q7Z3C6" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag> was</span> <sd-tag id="sdTag291" source="sdapi" category="assay" entity_type="" role="" text="immunoprecipitated" ext_ids="BAO_0002508" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunoprecipitation" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&amp;conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoprecipitated</sd-tag> <span>with an anti‐m<sd-tag id="sdTag559" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q674R7///Q7Z3C6" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag> antibody, the</span> <sd-tag id="sdTag560" source="sdapi" category="assay" entity_type="" role="" text="co‐immunoprecipitated" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">co‐immunoprecipitated</sd-tag> <sd-tag id="sdTag294" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag295" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38IP" ext_ids="Q8NEM7" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38IP</sd-tag> was detected with an anti‐<sd-tag id="sdTag296" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag> antibody, and <sd-tag id="sdTag561" source="sdapi" category="assay" entity_type="" role="" text="co‐immunoprecipitated" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">co‐immunoprecipitated</sd-tag> <sd-tag id="sdTag298" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag>-<sd-tag id="sdTag299" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38" ext_ids="O15264///P53778///Q15759///Q16539" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAPK13///MAPK12///MAPK11///MAPK14" ext_tax_ids="9606///9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">p38</sd-tag> was detected with an anti‐<sd-tag id="sdTag300" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag> antibody. <sd-tag id="sdTag301" source="sdapi" category="entity" entity_type="protein" role="reporter" text="RFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">RFP</sd-tag><span>-m<sd-tag id="sdTag562" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q674R7///Q7Z3C6" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag> was detected with an anti‐</span><sd-tag id="sdTag275" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q674R7///Q7Z3C6" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag> antibody.<graphic href="https://api.sourcedata.io/file.php?panel_id=4856"/></sd-panel><sd-panel panel_id="4857"><b>(B)</b> <sd-tag id="sdTag305" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HEK293A" ext_ids="CVCL_6910" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293-A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HEK293A</sd-tag> cells were transfected with <sd-tag id="sdTag306" source="sdapi" category="entity" entity_type="protein" role="reporter" text="RFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">RFP</sd-tag>-<span><sd-tag id="sdTag564" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q674R7///Q7Z3C6" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag></span> and <sd-tag id="sdTag308" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag565" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38IP" ext_ids="Q8NEM7" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38IP</sd-tag> (lanes 1 and 5), Flag-Flag, <sd-tag id="sdTag312" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag>-<span><sd-tag id="sdTag571" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38" ext_ids="O15264///P53778///Q15759///Q16539" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAPK13///MAPK12///MAPK11///MAPK14" ext_tax_ids="9606///9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">p38</sd-tag></span>, and <sd-tag id="sdTag314" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag566" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38IP" ext_ids="Q8NEM7" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38IP</sd-tag> (lanes 2 and 6), or <sd-tag id="sdTag316" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag>-<span><sd-tag id="sdTag570" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38" ext_ids="O15264///P53778///Q15759///Q16539" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAPK13///MAPK12///MAPK11///MAPK14" ext_tax_ids="9606///9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">p38</sd-tag></span> and <sd-tag id="sdTag318" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag567" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38IP" ext_ids="Q8NEM7" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38IP</sd-tag> (lanes 3 and 7). <sd-tag id="sdTag320" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag321" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38IP" ext_ids="Q8NEM7" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38IP</sd-tag> was <sd-tag id="sdTag322" source="sdapi" category="assay" entity_type="" role="" text="immunoprecipitated" ext_ids="BAO_0002508" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunoprecipitation" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&amp;conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoprecipitated</sd-tag> with an anti‐<sd-tag id="sdTag323" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag> antibody, the <sd-tag id="sdTag568" source="sdapi" category="assay" entity_type="" role="" text="co‐immunoprecipitated" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">co‐immunoprecipitated</sd-tag> <sd-tag id="sdTag325" source="sdapi" category="entity" entity_type="protein" role="reporter" text="RFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">RFP</sd-tag>-<sd-tag id="sdTag326" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q674R7///Q7Z3C6" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag> was detected with an anti‐<sd-tag id="sdTag327" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q674R7///Q7Z3C6" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag> antibody, and <sd-tag id="sdTag569" source="sdapi" category="assay" entity_type="" role="" text="co‐immunoprecipitated" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">co‐immunoprecipitated</sd-tag> <sd-tag id="sdTag329" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag>-<sd-tag id="sdTag330" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38" ext_ids="O15264///P53778///Q15759///Q16539" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAPK13///MAPK12///MAPK11///MAPK14" ext_tax_ids="9606///9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">p38</sd-tag> was detected with an anti‐<sd-tag id="sdTag304" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38α" ext_ids="Q16539" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="MAPK14" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38α</sd-tag> antibody. <sd-tag id="sdTag331" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag332" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38IP" ext_ids="Q8NEM7" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38IP</sd-tag> was detected with an anti‐<sd-tag id="sdTag333" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag> antibody.<graphic href="https://api.sourcedata.io/file.php?panel_id=4857"/></sd-panel><sd-panel panel_id="4858"><b>(C)</b> <sd-tag id="sdTag336" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HEK293A" ext_ids="CVCL_6910" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293-A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HEK293A</sd-tag> cells were transfected with <sd-tag id="sdTag337" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag>-<sd-tag id="sdTag335" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38α" ext_ids="1432" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="MAPK14" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38α</sd-tag> for 24 h. Cells were incubated in either full medium, <sd-tag id="sdTag339" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag>, or <sd-tag id="sdTag340" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> plus <sd-tag id="sdTag341" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="leupeptin" ext_ids="CHEBI:6426" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Leupeptin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">leupeptin</sd-tag> for 2 h, lysed, and analysed for endogenous <sd-tag id="sdTag343" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9BXW4///Q9H492///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3C///MAP1LC3A///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag> lipidation using an anti‐<sd-tag id="sdTag344" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9H492///Q9GZQ8///Q9BXW4" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3A///MAP1LC3B///MAP1LC3C" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag> antibody, and <sd-tag id="sdTag345" source="sdapi" category="assay" entity_type="" role="" text="immunoblotted" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&amp;conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotted</sd-tag> with anti‐<sd-tag id="sdTag346" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag> antibody. <sd-tag id="sdTag347" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9BXW4///Q9H492///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3C///MAP1LC3A///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag> lipidation was quantified as the amount of LC3II/LC3I (data are represented as mean±s.e.m. of triplicates, <i>n</i>=2 experiments, <sd-tag id="sdTag350" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> control versus <sd-tag id="sdTag351" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> with <sd-tag id="sdTag352" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag>‐<span><sd-tag id="sdTag572" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38α" ext_ids="Q16539" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="MAPK14" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38α</sd-tag></span>, <sup>***</sup><i>P</i>=0.0026).<graphic href="https://api.sourcedata.io/file.php?panel_id=4858"/></sd-panel></fig><fig id="1878"><title></title><label>Figure 7</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1878"/><sd-panel panel_id="4859"><b>(A)</b> <sd-tag id="sdTag354" source="sdapi" category="entity" entity_type="cell_line" role="component" text="293" ext_ids="CVCL_0045" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">293</sd-tag>/<sd-tag id="sdTag355" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag356" source="sdapi" category="entity" entity_type="gene" role="component" text="LC3" ext_ids="440738///81631///84557" norm_text="" ext_dbs="NCBI gene///NCBI gene///NCBI gene" in_caption="True" ext_names="MAP1LC3C///MAP1LC3B///MAP1LC3A" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene/">LC3</sd-tag> cells were transfected with control or <sd-tag id="sdTag498" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38α" ext_ids="5594" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="MAPK1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38α</sd-tag> siRNA. At <sd-pretag id="sdPretag14961398097980" parent-tag-id="590">72 h</sd-pretag> after transfection, cells were incubated in either full medium, <sd-tag id="sdTag360" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag>, or <sd-tag id="sdTag361" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> with <sd-tag id="sdTag362" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="leupeptin" ext_ids="CHEBI:6426" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Leupeptin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">leupeptin</sd-tag> for 2 h, then fixed and visualized by <sd-tag id="sdTag364" source="sdapi" category="assay" entity_type="" role="" text="confocal microscopy" ext_ids="BAO_0000453" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="confocal microscopy" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&amp;conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">confocal microscopy</sd-tag>. Bars=5 μm (data are represented as mean±s.e.m. <i>n</i>=60 cells, <sd-tag id="sdTag365" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> control versus <sd-tag id="sdTag499" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38α" ext_ids="5594" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="MAPK1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38α</sd-tag> siRNA (<sup>***</sup><i>P</i>=0.0001); <sd-tag id="sdTag368" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> with <sd-tag id="sdTag369" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="leupeptin" ext_ids="CHEBI:6426" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Leupeptin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">leupeptin</sd-tag> control versus <sd-tag id="sdTag500" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38α" ext_ids="5594" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="MAPK1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38α</sd-tag> siRNA (<sup>***</sup><i>P</i>=0.0001), Student's <i>t</i>‐test).<graphic href="https://api.sourcedata.io/file.php?panel_id=4859"/></sd-panel><sd-panel panel_id="4860"><b>(B)</b> <sd-tag id="sdTag379" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HEK293A" ext_ids="CVCL_6910" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293-A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HEK293A</sd-tag> cells were transfected with control or <sd-tag id="sdTag372" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38α" ext_ids="1432" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="MAPK14" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38α</sd-tag> siRNA. At <sd-pretag id="sdPretag14961397594840" parent-tag-id="590">72 h</sd-pretag> after transfection, cells were incubated in either full medium, <sd-tag id="sdTag381" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag>, or <sd-tag id="sdTag382" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> containing <sd-tag id="sdTag383" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="leupeptin" ext_ids="CHEBI:6426" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Leupeptin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">leupeptin</sd-tag> for 2 h. Cells lysates were analysed for <sd-tag id="sdTag373" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9H492///Q9BXW4///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3A///MAP1LC3C///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag> lipidation using an anti‐<sd-tag id="sdTag374" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9H492///Q9BXW4///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3A///MAP1LC3C///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag> antibody. The membrane was also probed with anti‐<sd-tag id="sdTag386" source="sdapi" category="entity" entity_type="protein" role="normalizing" text="actin" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">actin</sd-tag> and anti‐<sd-tag id="sdTag502" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38α" ext_ids="Q16539" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="MAPK14" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38α</sd-tag> antibodies. <sd-tag id="sdTag387" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9H492///Q9BXW4///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3A///MAP1LC3C///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag> lipidation was quantified as the amount of <span><sd-tag id="sdTag576" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9H492///Q9BXW4///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3A///MAP1LC3C///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag>II</span>/<span><sd-tag id="sdTag577" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9H492///Q9GZQ8///Q9BXW4" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3A///MAP1LC3B///MAP1LC3C" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag>I</span> (data are represented as mean ±s.e.m. <i>n</i>=5, full medium control versus <sd-tag id="sdTag376" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38α" ext_ids="1432" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="MAPK14" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38α</sd-tag> siRNA (<sup>**</sup><i>P</i>=0.0056); <sd-tag id="sdTag390" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> control versus <sd-tag id="sdTag377" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38α" ext_ids="1432" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="MAPK14" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38α</sd-tag> siRNA (<sup>**</sup><i>P</i>=0.048); <sd-tag id="sdTag391" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> with Leu control versus <sd-tag id="sdTag501" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38α" ext_ids="Q16539" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="MAPK14" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38α</sd-tag> siRNA (<sup>**</sup><i>P</i>=0.04), Student's <i>t</i>‐test).<graphic href="https://api.sourcedata.io/file.php?panel_id=4860"/></sd-panel><sd-panel panel_id="4861"><b>(C)</b> <sd-tag id="sdTag398" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HEK293A" ext_ids="CVCL_6910" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293-A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HEK293A</sd-tag> cells were transfected with control or <sd-tag id="sdTag581" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38α" ext_ids="1432" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="MAPK14" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38α</sd-tag> siRNA. At <sd-pretag id="sdPretag14961397995880" parent-tag-id="590">72 h</sd-pretag> after transfection, cells were incubated in either full medium or <sd-tag id="sdTag400" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> for 2 h, then fixed and <sd-tag id="sdTag402" source="sdapi" category="assay" entity_type="" role="" text="immunostained" ext_ids="BAO_0002426" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunofluorescent labeling" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&amp;conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunostained</sd-tag> to detect endogenous <span>m<sd-tag id="sdTag580" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q7Z3C6///Q674R7" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9A///ATG9B" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag></span> localization. The percentage of cells with dispersed <span>m<sd-tag id="sdTag579" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg9" ext_ids="Q674R7///Q7Z3C6" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG9B///ATG9A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Atg9</sd-tag></span> was quantified as in Figure 2E (data are represented as mean±s.e.m. <i>n</i>=200 cells, full medium control versus <sd-tag id="sdTag396" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38α" ext_ids="1432" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="MAPK14" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38α</sd-tag> siRNA (<sup>**</sup><i>P</i>=0.0027); <sd-tag id="sdTag404" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> control versus <sd-tag id="sdTag397" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38α" ext_ids="1432" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="MAPK14" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38α</sd-tag> siRNA (<sup>*</sup><i>P</i>=0.0459); Student's <i>t</i>‐test). Bars=5 μm<graphic href="https://api.sourcedata.io/file.php?panel_id=4861"/></sd-panel><sd-panel panel_id="4862"><b>(D)</b> <sd-tag id="sdTag409" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HEK293A" ext_ids="CVCL_6910" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293-A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HEK293A</sd-tag> cells were transfected with siRNA for <sd-tag id="sdTag503" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38α" ext_ids="1432" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="MAPK14" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38α</sd-tag>, homogenized and subjected to centrifugation, and the resulting post‐nuclear supernatant (PNS) was fractionated by centrifugation at 100 000 <i>g</i> into <sd-tag id="sdTag411" source="sdapi" category="entity" entity_type="subcellular" role="experiment" text="membrane" ext_ids="GO:0016020" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="membrane" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">membrane</sd-tag> pellet and <sd-tag id="sdTag412" source="sdapi" category="entity" entity_type="subcellular" role="experiment" text="cytosol" ext_ids="GO:0005829" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="cytosol" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">cytosol</sd-tag> as in Figure 2A. Equal protein amounts were resolved by <sd-tag id="sdTag413" source="sdapi" category="assay" entity_type="" role="" text="SDS–PAGE" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SDS-PAGE</sd-tag> and <sd-tag id="sdTag414" source="sdapi" category="assay" entity_type="" role="" text="immunoblotted" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&amp;conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotted</sd-tag> with anti‐<sd-tag id="sdTag415" source="sdapi" category="entity" entity_type="protein" role="assayed" text="MPR" ext_ids="P20645" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="M6PR" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">MPR</sd-tag>, anti‐<sd-tag id="sdTag416" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>, and anti‐<sd-tag id="sdTag407" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38α" ext_ids="Q16539" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="MAPK14" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38α</sd-tag> antibodies. Data are representative of four experiments.<graphic href="https://api.sourcedata.io/file.php?panel_id=4862"/></sd-panel><sd-panel panel_id="4863"><p><strong>(E)</strong> <sd-tag id="sdTag590">72 h</sd-tag> cells were transfected with control, <sd-tag id="sdTag419" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38α" ext_ids="1432" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="MAPK14" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38α</sd-tag> siRNA or both <sd-tag id="sdTag420" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38α" ext_ids="1432" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="MAPK14" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38α</sd-tag> siRNA, and <sd-tag id="sdTag421" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p38IP" ext_ids="55578" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="SUPT20H" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p38IP</sd-tag> siRNA. At <sd-pretag id="sdPretag14961398430410" parent-tag-id="590">72 h</sd-pretag> after transfection, cells were incubated in either full medium, full medium with <sd-tag id="sdTag428" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="leupeptin" ext_ids="CHEBI:6426" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Leupeptin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">leupeptin</sd-tag>, <sd-tag id="sdTag429" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag>, or <sd-tag id="sdTag430" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> with <sd-tag id="sdTag431" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="leupeptin" ext_ids="CHEBI:6426" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Leupeptin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">leupeptin</sd-tag> for 2 h. Cells lysates were analysed for <sd-tag id="sdTag583" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9BXW4///Q9H492///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3C///MAP1LC3A///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag> lipidation using an anti‐<sd-tag id="sdTag423" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9BXW4///Q9H492///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3C///MAP1LC3A///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag> antibody. The membrane was also probed with anti‐<sd-tag id="sdTag424" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p38α" ext_ids="Q16539" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="MAPK14" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">p38α</sd-tag>. <sd-tag id="sdTag425" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9BXW4///Q9H492///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3C///MAP1LC3A///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag> lipidation was quantified as the amount of LC3II/LC3I. Data are representative of two experiments.</p><graphic href="https://api.sourcedata.io/file.php?panel_id=4863"/></sd-panel></fig><fig id="1879"><title></title><label>Figure 8</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1879"/></fig></article>