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<article doi="10.1002/emmm.201303356" abstract="In addition to key roles in embryonic neurogenesis and myelinogenesis, γ-aminobutyric acid (GABA) serves as the primary inhibitory mammalian neurotransmitter. In yeast, we have identified a new role for GABA that augments activity of the pivotal kinase, Tor1. GABA inhibits the selective autophagy pathways, mitophagy and pexophagy, through Sch9, the homolog of the mammalian kinase, S6K1, leading to oxidative stress, all of which can be mitigated by the Tor1 inhibitor, rapamycin. To confirm these processes in mammals, we examined the succinic semialdehyde dehydrogenase (SSADH)-deficient mouse model that accumulates supraphysiological GABA in the central nervous system and other tissues. Mutant mice displayed increased mitochondrial numbers in the brain and liver, expected with a defect in mitophagy, and morphologically abnormal mitochondria. Administration of rapamycin to these mice reduced mTOR activity, reduced the elevated mitochondrial numbers, and normalized aberrant antioxidant levels. These results confirm a novel role for GABA in cell signaling and highlight potential pathomechanisms and treatments in various human pathologies, including SSADH deficiency, as well as other diseases characterized by elevated levels of GABA."><fig id="1152"><title></title><label>Figure 1</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1152"/><sd-panel panel_id="2386"><div>A <sd-tag id="sdTag1" source="sdapi" category="entity" entity_type="subcellular" role="component" text="Peroxisomes" ext_ids="GO:0005777" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="peroxisome" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">Peroxisomes</sd-tag> were induced by growing the WT strain expressing <sd-tag id="sdTag3" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Pot1" ext_ids="P36002" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PTK1" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Pot1</sd-tag>‐<sd-tag id="sdTag4" 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> in <sd-tag id="sdTag5" source="sdapi" category="entity" entity_type="undefined" role="component" text="oleate medium" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">oleate medium</sd-tag> to mid‐log‐phase, then transferred to <sd-tag id="sdTag380" source="sdapi" category="entity" entity_type="undefined" role="component" text="SD‐N" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SD‐N</sd-tag> starvation medium with or without <sd-tag id="sdTag2" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="GABA" ext_ids="CHEBI:16865" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="gamma-aminobutyric acid" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">GABA</sd-tag> to trigger pexophagy for 6 h. <sd-tag id="sdTag8" 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> cleavage was analyzed at the indicated time points by <sd-tag id="sdTag7" source="sdapi" category="assay" entity_type="" role="" text="immunoblotting" 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&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotting</sd-tag>.</div><graphic href="https://api.sourcedata.io/file.php?panel_id=2386"/></sd-panel><sd-panel panel_id="2387">B <sd-tag id="sdTag18" source="sdapi" category="entity" entity_type="subcellular" role="component" text="Mitochondria" ext_ids="GO:0005739" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="mitochondrion" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">Mitochondria</sd-tag> were induced by growing the WT strain expressing <sd-tag id="sdTag14" source="sdapi" category="entity" entity_type="protein" role="assayed" text="OM45" ext_ids="P16547" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="OM45" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">OM45</sd-tag>‐<sd-tag id="sdTag10" 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> in <sd-tag id="sdTag36" source="sdapi" category="entity" entity_type="undefined" role="component" text="YPL medium" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">YPL medium</sd-tag> to mid‐log‐phase and subsequently transferring cells to either <sd-tag id="sdTag16" source="sdapi" category="entity" entity_type="undefined" role="component" text="SD‐N" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SD‐N</sd-tag> with or without <sd-tag id="sdTag11" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="GABA" ext_ids="CHEBI:16865" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="gamma-aminobutyric acid" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">GABA</sd-tag> to trigger mitophagy for 12 h. <sd-tag id="sdTag12" 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> cleavage was analyzed at the indicated time points by <sd-tag id="sdTag13" source="sdapi" category="assay" entity_type="" role="" text="immunoblotting" 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&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotting</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=2387"/></sd-panel><sd-panel panel_id="2388">C Mitophagy was monitored by <sd-tag id="sdTag20" source="sdapi" category="assay" entity_type="" role="" text="fluorescence microscopy" ext_ids="BAO_0000450" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="fluorescence microscopy" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">fluorescence microscopy</sd-tag> using a WT strain expressing <sd-tag id="sdTag21" source="sdapi" category="entity" entity_type="protein" role="assayed" text="OM45" ext_ids="P16547" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="OM45" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">OM45</sd-tag>‐<sd-tag id="sdTag22" 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> grown in <sd-tag id="sdTag25" source="sdapi" category="entity" entity_type="undefined" role="component" text="YPL medium" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">YPL medium</sd-tag> for 12 h to mid‐log‐phase in the presence of <sd-tag id="sdTag26" source="sdapi" category="assay" entity_type="" role="" text="FM4‐64" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">FM4‐64</sd-tag>, and transferred to either <sd-tag id="sdTag381" source="sdapi" category="entity" entity_type="undefined" role="component" text="SD‐N" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SD‐N</sd-tag> medium with or without <sd-tag id="sdTag24" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="GABA" ext_ids="CHEBI:16865" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="gamma-aminobutyric acid" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">GABA</sd-tag> for 24 h. Bar, 5 μm.<graphic href="https://api.sourcedata.io/file.php?panel_id=2388"/></sd-panel><sd-panel panel_id="2389">D The Cvt pathway was monitored using the WT strain in <sd-tag id="sdTag34" source="sdapi" category="entity" entity_type="undefined" role="component" text="SD medium" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SD medium</sd-tag> with or without <sd-tag id="sdTag30" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="GABA" ext_ids="CHEBI:16865" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="gamma-aminobutyric acid" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">GABA</sd-tag>, grown to mid‐log‐phase, after which samples were analyzed for <sd-tag id="sdTag31" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Ape1" ext_ids="P14904" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="APE1" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Ape1</sd-tag> maturation.<graphic href="https://api.sourcedata.io/file.php?panel_id=2389"/></sd-panel><sd-panel panel_id="2390">E Ribophagy was monitored by growing the WT strain expressing <sd-tag id="sdTag41" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Rpl25" ext_ids="P04456" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="RPL25" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Rpl25</sd-tag>‐<sd-tag id="sdTag37" 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> in <sd-tag id="sdTag38" source="sdapi" category="entity" entity_type="undefined" role="component" text="SD medium" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SD medium</sd-tag> to mid‐log‐phase and transferring cells to <sd-tag id="sdTag39" source="sdapi" category="entity" entity_type="undefined" role="component" text="SD‐N" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SD‐N</sd-tag> either with or without <sd-tag id="sdTag40" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="GABA" ext_ids="CHEBI:16865" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="gamma-aminobutyric acid" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">GABA</sd-tag> for 24 h.<graphic href="https://api.sourcedata.io/file.php?panel_id=2390"/></sd-panel><sd-panel panel_id="2391">F Autophagy was monitored by growing the WT strain expressing <sd-tag id="sdTag44" 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="sdTag48" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Atg8</sd-tag> in <sd-tag id="sdTag45" source="sdapi" category="entity" entity_type="undefined" role="component" text="SD medium" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SD medium</sd-tag> to mid‐log‐phase and transferring cells to <sd-tag id="sdTag46" source="sdapi" category="entity" entity_type="undefined" role="component" text="SD‐N" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SD‐N</sd-tag> either with or without <sd-tag id="sdTag47" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="GABA" ext_ids="CHEBI:16865" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="gamma-aminobutyric acid" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">GABA</sd-tag> for 6 h.<graphic href="https://api.sourcedata.io/file.php?panel_id=2391"/></sd-panel></fig><fig id="1153"><title></title><label>Figure 2</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1153"/><sd-panel panel_id="2392">A<sd-tag id="sdTag51" source="sdapi" category="entity" entity_type="subcellular" role="component" text="Peroxisomes" ext_ids="GO:0005777" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="peroxisome" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">Peroxisomes</sd-tag> were induced in <sd-tag id="sdTag52" source="sdapi" category="entity" entity_type="undefined" role="component" text="oleate medium" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">oleate medium</sd-tag> and pexophagy was monitored as described for Fig 1A.<graphic href="https://api.sourcedata.io/file.php?panel_id=2392"/></sd-panel><sd-panel panel_id="2393">B Pexophagy was monitored by <sd-tag id="sdTag63" source="sdapi" category="assay" entity_type="" role="" text="fluorescence microscopy" ext_ids="BAO_0000450" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="fluorescence microscopy" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">fluorescence microscopy</sd-tag> using a WT strain expressing <sd-tag id="sdTag64" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Pot1" ext_ids="P36002" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PTK1" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Pot1</sd-tag>‐<sd-tag id="sdTag65" 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> grown in <sd-tag id="sdTag66" source="sdapi" category="entity" entity_type="undefined" role="component" text="oleate medium" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">oleate medium</sd-tag> to mid‐log‐phase in the presence of <sd-tag id="sdTag67" source="sdapi" category="assay" entity_type="" role="" text="FM4‐64" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">FM4‐64</sd-tag>, and transferred to either <sd-tag id="sdTag68" source="sdapi" category="entity" entity_type="undefined" role="component" text="SD‐N" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SD‐N</sd-tag> medium with or without <sd-tag id="sdTag69" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="GABA" ext_ids="CHEBI:16865" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="gamma-aminobutyric acid" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">GABA</sd-tag> or to <sd-tag id="sdTag70" source="sdapi" category="entity" entity_type="undefined" role="component" text="SD‐N" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SD‐N</sd-tag> with <sd-tag id="sdTag71" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="GABA" ext_ids="CHEBI:16865" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="gamma-aminobutyric acid" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">GABA</sd-tag> and <sd-tag id="sdTag72" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="rapamycin" ext_ids="CHEBI:9168" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="sirolimus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">rapamycin</sd-tag> for 6 h. Bar, 5 μm.<graphic href="https://api.sourcedata.io/file.php?panel_id=2393"/></sd-panel><sd-panel panel_id="2394">C <sd-tag id="sdTag75" source="sdapi" category="entity" entity_type="subcellular" role="component" text="Mitochondria" ext_ids="GO:0005739" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="mitochondrion" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">Mitochondria</sd-tag> were induced in <sd-tag id="sdTag76" source="sdapi" category="entity" entity_type="undefined" role="component" text="YPL medium" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">YPL medium</sd-tag> and mitophagy was assessed as described for Fig .<graphic href="https://api.sourcedata.io/file.php?panel_id=2394"/></sd-panel></fig><fig id="1154"><title></title><label>Figure 3</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1154"/><sd-panel panel_id="2395">A WT cells expressing <sd-tag id="sdTag107" source="sdapi" category="entity" entity_type="protein" role="assayed" text="OM45" ext_ids="P16547" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="OM45" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">OM45</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>, along with the <sd-tag id="sdTag365" source="sdapi" category="entity" entity_type="gene" role="intervention" text="uga2" ext_ids="852291" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="UGA2" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">uga2</sd-tag>∆ strain over‐expressing the <sd-tag id="sdTag116" source="sdapi" category="entity" entity_type="gene" role="intervention" text="GAD1" ext_ids="855291" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="GAD1" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">GAD1</sd-tag> gene and expressing <sd-tag id="sdTag108" source="sdapi" category="entity" entity_type="protein" role="assayed" text="OM45" ext_ids="P16547" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="OM45" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">OM45</sd-tag>‐<sd-tag id="sdTag110" 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> were grown in <sd-tag id="sdTag111" source="sdapi" category="entity" entity_type="undefined" role="component" text="YPL medium" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">YPL medium</sd-tag> to mid‐log‐phase. To monitor mitophagy, strains were transferred to SD‐N starvation medium (with or without <sd-tag id="sdTag114" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="rapamycin" ext_ids="CHEBI:9168" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="sirolimus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">rapamycin</sd-tag>).<graphic href="https://api.sourcedata.io/file.php?panel_id=2395"/></sd-panel><sd-panel panel_id="2396">B WT strain along with the <sd-tag id="sdTag366" source="sdapi" category="entity" entity_type="gene" role="intervention" text="uga2" ext_ids="852291" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="UGA2" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">uga2</sd-tag>∆ strain over‐expressing the <sd-tag id="sdTag119" source="sdapi" category="entity" entity_type="gene" role="intervention" text="GAD1" ext_ids="855291" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="GAD1" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">GAD1</sd-tag> gene was grown in <sd-tag id="sdTag120" source="sdapi" category="entity" entity_type="undefined" role="component" text="oleate medium" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">oleate medium</sd-tag> and pexophagy was monitored as described in Fig , with or without <sd-tag id="sdTag121" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="rapamycin" ext_ids="CHEBI:9168" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="sirolimus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">rapamycin</sd-tag>. Samples were taken at the indicated time points, and <sd-tag id="sdTag122" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Pot1" ext_ids="P36002" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PTK1" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Pot1</sd-tag> degradation was analyzed by <sd-tag id="sdTag123" source="sdapi" category="assay" entity_type="" role="" text="immunoblotting" 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&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotting</sd-tag> (45 kD).<graphic href="https://api.sourcedata.io/file.php?panel_id=2396"/></sd-panel><sd-panel panel_id="2397">C To monitor autophagy, WT cells expressing <sd-tag id="sdTag126" 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="sdTag127" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Atg8</sd-tag> along with the <sd-tag id="sdTag368" source="sdapi" category="entity" entity_type="gene" role="intervention" text="uga2" ext_ids="852291" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="UGA2" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">uga2</sd-tag>∆ strain over‐expressing the <sd-tag id="sdTag129" source="sdapi" category="entity" entity_type="gene" role="intervention" text="GAD1" ext_ids="855291" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="GAD1" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">GAD1</sd-tag> gene and expressing <sd-tag id="sdTag130" 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="sdTag131" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Atg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Atg8</sd-tag> were grown in <sd-tag id="sdTag132" source="sdapi" category="entity" entity_type="undefined" role="component" text="SD medium" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SD medium</sd-tag> and transferred to <sd-tag id="sdTag133" source="sdapi" category="entity" entity_type="undefined" role="component" text="SD‐N" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SD‐N</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=2397"/></sd-panel></fig><fig id="1155"><title></title><label>Figure 4</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1155"/><sd-panel panel_id="2398">A,B WT cells were cultured under pexophagy (A) or mitophagy (B) conditions with or without <sd-tag id="sdTag135" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="GABA" ext_ids="CHEBI:16865" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="gamma-aminobutyric acid" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">GABA</sd-tag> and <sd-tag id="sdTag136" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="rapamycin" ext_ids="CHEBI:9168" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="sirolimus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">rapamycin</sd-tag>. <sd-tag id="sdTag138" source="sdapi" category="entity" entity_type="protein" role="assayed" text="S6" ext_ids="P0CX37///P0CX38" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="RPS6A///RPS6B" ext_tax_ids="559292///559292" ext_tax_names="Saccharomyces cerevisiae S288C///Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">S6</sd-tag> phosphorylation at the indicated time points was analyzed by <sd-tag id="sdTag137" source="sdapi" category="assay" entity_type="" role="" text="immunoblotting" 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&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotting</sd-tag> with a loading control.<graphic href="https://api.sourcedata.io/file.php?panel_id=2398"/></sd-panel><sd-panel panel_id="2399">C Samples were analyzed for <sd-tag id="sdTag140" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Pot1" ext_ids="P36002" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PTK1" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Pot1</sd-tag> degradation by <sd-tag id="sdTag141" source="sdapi" category="assay" entity_type="" role="" text="immunoblotting" 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&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotting</sd-tag> (45 kD).<graphic href="https://api.sourcedata.io/file.php?panel_id=2399"/></sd-panel><sd-panel panel_id="2400">D <sd-tag id="sdTag144" 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> production during mitophagy was analyzed by <sd-tag id="sdTag145" source="sdapi" category="assay" entity_type="" role="" text="immunoblotting" 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&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotting</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=2400"/></sd-panel></fig><fig id="1156"><title></title><label>Figure 5</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1156"/><sd-panel panel_id="2402">A <sd-tag id="sdTag150" source="sdapi" category="entity" entity_type="protein" role="assayed" text="S6" ext_ids="P0CX37///P0CX38" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="RPS6A///RPS6B" ext_tax_ids="559292///559292" ext_tax_names="Saccharomyces cerevisiae S288C///Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">S6</sd-tag> phosphorylation after 6 h in <sd-tag id="sdTag152" source="sdapi" category="entity" entity_type="undefined" role="component" text="SD‐N" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SD‐N</sd-tag> was analyzed by <sd-tag id="sdTag151" source="sdapi" category="assay" entity_type="" role="" text="immunoblotting" 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&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotting</sd-tag> with a loading control.<graphic href="https://api.sourcedata.io/file.php?panel_id=2402"/></sd-panel><sd-panel panel_id="2403">B <sd-tag id="sdTag157" 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> production monitoring autophagy at the indicated time points was analyzed by <sd-tag id="sdTag158" source="sdapi" category="assay" entity_type="" role="" text="immunoblotting" 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&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotting</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=2403"/></sd-panel></fig><fig id="1157"><title></title><label>Figure 6</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1157"/><sd-panel panel_id="2404">A,B WT, WT with <sd-tag id="sdTag162" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="GABA" ext_ids="CHEBI:16865" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="gamma-aminobutyric acid" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">GABA</sd-tag>, WT with <sd-tag id="sdTag163" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="GABA" ext_ids="CHEBI:16865" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="gamma-aminobutyric acid" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">GABA</sd-tag> and 10 mM <sd-tag id="sdTag166" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="GSH" ext_ids="CHEBI:16856" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="glutathione" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">GSH</sd-tag> and WT with <sd-tag id="sdTag164" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="GABA" ext_ids="CHEBI:16865" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="gamma-aminobutyric acid" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">GABA</sd-tag> and 200 nM <sd-tag id="sdTag165" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="rapamycin" ext_ids="CHEBI:9168" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="sirolimus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">rapamycin</sd-tag> were tested for <sd-tag id="sdTag172" source="sdapi" category="entity" entity_type="molecule" role="assayed" text="intracellular ROS" ext_ids="CHEBI:26523" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="reactive oxygen species" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">intracellular ROS</sd-tag> levels under (A) pexophagy and (B) mitophagy conditions. After 24 h incubation, cells were stained with <sd-tag id="sdTag338" source="sdapi" category="assay" entity_type="" role="" text="DHR−123" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">DHR−123</sd-tag> and <sd-tag id="sdTag168" source="sdapi" category="assay" entity_type="" role="" text="propidium iodide" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">propidium iodide</sd-tag> for 1 h. Living cells were analyzed for <sd-tag id="sdTag169" source="sdapi" category="assay" entity_type="" role="" text="DHR−123" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">DHR−123</sd-tag> fluorescence by <sd-tag id="sdTag170" source="sdapi" category="assay" entity_type="" role="" text="flow cytometry" ext_ids="BAO_0000005" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="flow cytometry" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">flow cytometry</sd-tag>. Data represent mean + s.d. (n = 3). *P < 0.005, **P < 0.01<graphic href="https://api.sourcedata.io/file.php?panel_id=2404"/></sd-panel><sd-panel panel_id="2405">C,D <sd-tag id="sdTag175" source="sdapi" category="entity" entity_type="organism" role="assayed" text="Yeast" 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/">Yeast</sd-tag> cells stained with 5 μM <sd-tag id="sdTag174" source="sdapi" category="assay" entity_type="" role="" text="propidium iodide" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">propidium iodide</sd-tag> were used to differentiate between living and dead cells under (C) pexophagy or (D) mitophagy conditions. Significant differences between the treatments and strains were determined using an unpaired two‐tailed t‐test. **P < 0.01.<graphic href="https://api.sourcedata.io/file.php?panel_id=2405"/></sd-panel><sd-panel panel_id="2406">E Pexophagy assay was monitored by the degradation of <sd-tag id="sdTag178" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Pot1" ext_ids="P36002" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PTK1" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Pot1</sd-tag>‐<sd-tag id="sdTag179" 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> and analyzed for <sd-tag id="sdTag180" 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> cleavage by <sd-tag id="sdTag181" source="sdapi" category="assay" entity_type="" role="" text="immunoblotting" 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&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotting</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=2406"/></sd-panel><sd-panel panel_id="2407">F Mitophagy assay was monitored by the degradation of <sd-tag id="sdTag189" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Om45" ext_ids="P16547" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="OM45" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Om45</sd-tag>‐<sd-tag id="sdTag186" 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> and analyzed for <sd-tag id="sdTag187" 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> cleavage by <sd-tag id="sdTag188" source="sdapi" category="assay" entity_type="" role="" text="immunoblotting" 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&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotting</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=2407"/></sd-panel></fig><fig id="1158"><title></title><label>Figure 7</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1158"/><sd-panel panel_id="2408">A,B Example images of <sd-tag id="sdTag194" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Parkin" ext_ids="5071" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Parkin</sd-tag>‐expressing <sd-tag id="sdTag199" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HeLa cells" 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 cells</sd-tag> analyzed using a tandem fluorochrome protein (mito‐<sd-tag id="sdTag196" 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="sdTag195" 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>) mitophagy assay under (A) control conditions or (B) displaying mitophagy depicted by the red mitochondrial structures localized to <sd-tag id="sdTag198" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="lysosomes" ext_ids="GO:0005764" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="lysosome" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">lysosomes</sd-tag>. Bar, 10 μm.<graphic href="https://api.sourcedata.io/file.php?panel_id=2408"/></sd-panel><sd-panel panel_id="2409">C Percentage of cells displaying mitophagy + s.d., **P < 0.01 using an unpaired two‐tailed t‐test, n > 80.<graphic href="https://api.sourcedata.io/file.php?panel_id=2409"/></sd-panel></fig><fig id="1159"><title></title><label>Figure 8</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1159"/><sd-panel panel_id="2410"><div>A <sd-tag id="sdTag210" source="sdapi" category="assay" entity_type="" role="" text="Electron microscopy" ext_ids="BAO_0000449" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="electron microscopy" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">Electron microscopy</sd-tag> images of <sd-tag id="sdTag211" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="mitochondria" ext_ids="GO:0005739" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="mitochondrion" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">mitochondria</sd-tag> from WT (n = 44) and <sd-tag id="sdTag212" source="sdapi" category="entity" entity_type="protein" role="component" text="SSADH" ext_ids="Q8BWF0" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="Aldh5a1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="https://www.uniprot.org/uniprot/">SSADH</sd-tag>‐deficient <sd-tag id="sdTag300" source="sdapi" category="entity" entity_type="organism" role="component" text="mice" ext_ids="10090" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Mus musculus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">mice</sd-tag> (<sd-tag id="sdTag213" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Aldh5a1" ext_ids="214579" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Aldh5a1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Aldh5a1</sd-tag>−/−) (n = 80) were calculated for area size.</div><graphic href="https://api.sourcedata.io/file.php?panel_id=2410"/></sd-panel><sd-panel panel_id="2411">B <sd-tag id="sdTag214" source="sdapi" category="assay" entity_type="" role="" text="Electron microscopy" ext_ids="BAO_0000449" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="electron microscopy" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">Electron microscopy</sd-tag> images showing typical sizes of WT and <sd-tag id="sdTag215" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Aldh5a1" ext_ids="214579" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Aldh5a1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Aldh5a1</sd-tag>−/− <sd-tag id="sdTag218" source="sdapi" category="entity" entity_type="organism" role="component" text="mice" ext_ids="10090" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Mus musculus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">mice</sd-tag> <sd-tag id="sdTag217" source="sdapi" category="entity" entity_type="tissue" role="component" text="liver" ext_ids="UBERON:0002107" norm_text="" ext_dbs="Uberon" in_caption="True" ext_names="liver" ext_tax_ids="" ext_tax_names="" ext_urls="https://www.ebi.ac.uk/ols/ontologies/uberon/terms?iri=http%3A%2F%2Fpurl.obolibrary.org%2Fobo%2FUBERON_">liver</sd-tag> <sd-tag id="sdTag216" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="mitochondria" ext_ids="GO:0005739" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="mitochondrion" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">mitochondria</sd-tag>. Bar, 0.5 μm.<graphic href="https://api.sourcedata.io/file.php?panel_id=2411"/></sd-panel><sd-panel panel_id="2412">C Quantification of <sd-tag id="sdTag224" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="mitochondria" ext_ids="GO:0005739" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="mitochondrion" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">mitochondria</sd-tag>l numbers from <sd-tag id="sdTag223" source="sdapi" category="assay" entity_type="" role="" text="electron microscopy" ext_ids="BAO_0000449" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="electron microscopy" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">electron microscopy</sd-tag> images of <sd-tag id="sdTag219" source="sdapi" category="entity" entity_type="tissue" role="component" text="liver" ext_ids="UBERON:0002107" norm_text="" ext_dbs="Uberon" in_caption="True" ext_names="liver" ext_tax_ids="" ext_tax_names="" ext_urls="https://www.ebi.ac.uk/ols/ontologies/uberon/terms?iri=http%3A%2F%2Fpurl.obolibrary.org%2Fobo%2FUBERON_">liver</sd-tag> from WT (n = 31) and <sd-tag id="sdTag220" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Aldh5a1" ext_ids="214579" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Aldh5a1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Aldh5a1</sd-tag>−/− <sd-tag id="sdTag221" source="sdapi" category="entity" entity_type="organism" role="component" text="mice" ext_ids="10090" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Mus musculus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">mice</sd-tag> treated with vehicle (n = 39) or <sd-tag id="sdTag222" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="rapamycin" ext_ids="CHEBI:9168" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="sirolimus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">rapamycin</sd-tag> (n = 34) (5 mg/kg body weight per day) via intraperitoneal injections for 3 successive days starting at day 7 of life.<graphic href="https://api.sourcedata.io/file.php?panel_id=2412"/></sd-panel><sd-panel panel_id="2413">D Quantification of <sd-tag id="sdTag230" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="mitochondria" ext_ids="GO:0005739" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="mitochondrion" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">mitochondria</sd-tag>l numbers from <sd-tag id="sdTag225" source="sdapi" category="assay" entity_type="" role="" text="electron microscopy" ext_ids="BAO_0000449" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="electron microscopy" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">electron microscopy</sd-tag> images of <sd-tag id="sdTag229" source="sdapi" category="entity" entity_type="tissue" role="component" text="brain" ext_ids="UBERON:0000955" norm_text="" ext_dbs="Uberon" in_caption="True" ext_names="brain" ext_tax_ids="" ext_tax_names="" ext_urls="https://www.ebi.ac.uk/ols/ontologies/uberon/terms?iri=http%3A%2F%2Fpurl.obolibrary.org%2Fobo%2FUBERON_">brain</sd-tag> from WT (n = 23) and <sd-tag id="sdTag226" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Aldh5a1" ext_ids="214579" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Aldh5a1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Aldh5a1</sd-tag>−/− <sd-tag id="sdTag227" source="sdapi" category="entity" entity_type="organism" role="component" text="mice" ext_ids="10090" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Mus musculus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">mice</sd-tag> treated with vehicle (n = 30) or <sd-tag id="sdTag228" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="rapamycin" ext_ids="CHEBI:9168" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="sirolimus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">rapamycin</sd-tag> (n = 41) (5 mg/kg body weight per day) via intraperitoneal injections for 3 successive days starting at day 7 of life.<graphic href="https://api.sourcedata.io/file.php?panel_id=2413"/></sd-panel><sd-panel panel_id="2414">E <sd-tag id="sdTag231" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Aldh5a1" ext_ids="214579" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Aldh5a1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Aldh5a1</sd-tag>−/− <sd-tag id="sdTag232" source="sdapi" category="entity" entity_type="organism" role="component" text="mice" ext_ids="10090" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Mus musculus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">mice</sd-tag> were treated with vehicle or <sd-tag id="sdTag233" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="rapamycin" ext_ids="CHEBI:9168" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="sirolimus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">rapamycin</sd-tag> (10 mg/kg body weight per day) via intraperitoneal injections for 10 successive days starting at day 10 of life. WT <sd-tag id="sdTag234" source="sdapi" category="entity" entity_type="organism" role="component" text="mice" ext_ids="10090" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Mus musculus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">mice</sd-tag> served as non‐disease controls (set to 1). After sacrifice, <sd-tag id="sdTag235" source="sdapi" category="entity" entity_type="tissue" role="component" text="liver" ext_ids="UBERON:0002107" norm_text="" ext_dbs="Uberon" in_caption="True" ext_names="liver" ext_tax_ids="" ext_tax_names="" ext_urls="https://www.ebi.ac.uk/ols/ontologies/uberon/terms?iri=http%3A%2F%2Fpurl.obolibrary.org%2Fobo%2FUBERON_">liver</sd-tag> homogenates were used to measure <sd-tag id="sdTag236" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="SOD enzyme" ext_ids="GO:1902693" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="superoxide dismutase complex" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">SOD enzyme</sd-tag> activity using a <sd-tag id="sdTag316" source="sdapi" category="assay" entity_type="" role="" text="colorimetric SOD activity assay" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">colorimetric SOD activity assay</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=2414"/></sd-panel><sd-panel panel_id="2415">F <sd-tag id="sdTag240" source="sdapi" category="entity" entity_type="subcellular" role="component" text="Mitochondria" ext_ids="GO:0005739" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="mitochondrion" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">Mitochondria</sd-tag>l <sd-tag id="sdTag237" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SOD2" ext_ids="P09671" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="Sod2" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="https://www.uniprot.org/uniprot/">SOD2</sd-tag> protein levels were quantified from <sd-tag id="sdTag239" source="sdapi" category="entity" entity_type="tissue" role="component" text="liver" ext_ids="UBERON:0002107" norm_text="" ext_dbs="Uberon" in_caption="True" ext_names="liver" ext_tax_ids="" ext_tax_names="" ext_urls="https://www.ebi.ac.uk/ols/ontologies/uberon/terms?iri=http%3A%2F%2Fpurl.obolibrary.org%2Fobo%2FUBERON_">liver</sd-tag> microsections using <sd-tag id="sdTag238" source="sdapi" category="assay" entity_type="" role="" text="immunofluorescence microscopy" ext_ids="BAO_0000450" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="fluorescence microscopy" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunofluorescence microscopy</sd-tag> and automated image analysis (WT set to 1).<graphic href="https://api.sourcedata.io/file.php?panel_id=2415"/></sd-panel><sd-panel panel_id="2416">G <sd-tag id="sdTag250" source="sdapi" category="assay" entity_type="" role="" text="Immunofluorescence images" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Immunofluorescence images</sd-tag> showing typical <sd-tag id="sdTag251" source="sdapi" category="entity" entity_type="subcellular" role="component" text="nuclear" 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/">nuclear</sd-tag> staining (<sd-tag id="sdTag252" source="sdapi" category="assay" entity_type="" role="" text="DAPI" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">DAPI</sd-tag>, blue) and <sd-tag id="sdTag244" source="sdapi" category="entity" entity_type="protein" role="assayed" text="SOD2" ext_ids="P09671" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="Sod2" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="https://www.uniprot.org/uniprot/">SOD2</sd-tag> staining (red) from WT, <sd-tag id="sdTag245" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Aldh5a1" ext_ids="214579" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Aldh5a1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Aldh5a1</sd-tag>−/− <sd-tag id="sdTag246" source="sdapi" category="entity" entity_type="organism" role="component" text="mice" ext_ids="10090" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Mus musculus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">mice</sd-tag> treated with vehicle and <sd-tag id="sdTag247" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Aldh5a1" ext_ids="214579" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Aldh5a1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Aldh5a1</sd-tag>−/− <sd-tag id="sdTag248" source="sdapi" category="entity" entity_type="organism" role="component" text="mice" ext_ids="10090" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Mus musculus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">mice</sd-tag> treated with <sd-tag id="sdTag249" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="rapamycin" ext_ids="CHEBI:9168" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="sirolimus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">rapamycin</sd-tag>. Bar, 10 μm.<graphic href="https://api.sourcedata.io/file.php?panel_id=2416"/></sd-panel></fig><fig id="1160"><title></title><label>Figure 9</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1160"/><sd-panel panel_id="2417">A Quantification of <sd-tag id="sdTag253" source="sdapi" category="entity" entity_type="protein" role="assayed" text="S6" ext_ids="P62754" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="Rps6" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="https://www.uniprot.org/uniprot/">S6</sd-tag> phosphorylation of <sd-tag id="sdTag255" source="sdapi" category="entity" entity_type="tissue" role="component" text="liver" ext_ids="UBERON:0002107" norm_text="" ext_dbs="Uberon" in_caption="True" ext_names="liver" ext_tax_ids="" ext_tax_names="" ext_urls="https://www.ebi.ac.uk/ols/ontologies/uberon/terms?iri=http%3A%2F%2Fpurl.obolibrary.org%2Fobo%2FUBERON_">liver</sd-tag> lysates from WT (n = 5) and <sd-tag id="sdTag256" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Aldh5a1" ext_ids="214579" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Aldh5a1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Aldh5a1</sd-tag>−/− <sd-tag id="sdTag257" source="sdapi" category="entity" entity_type="organism" role="component" text="mice" ext_ids="10090" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Mus musculus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">mice</sd-tag> treated with vehicle (n = 4) or <sd-tag id="sdTag258" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="rapamycin" ext_ids="CHEBI:9168" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="sirolimus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">rapamycin</sd-tag> (n = 5) after normalization (WT set to 1).<graphic href="https://api.sourcedata.io/file.php?panel_id=2417"/></sd-panel><sd-panel panel_id="2418">B <sd-tag id="sdTag259" source="sdapi" category="entity" entity_type="protein" role="assayed" text="S6" ext_ids="P62754" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="Rps6" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="https://www.uniprot.org/uniprot/">S6</sd-tag> phosphorylation of <sd-tag id="sdTag261" source="sdapi" category="entity" entity_type="tissue" role="component" text="liver" ext_ids="UBERON:0002107" norm_text="" ext_dbs="Uberon" in_caption="True" ext_names="liver" ext_tax_ids="" ext_tax_names="" ext_urls="https://www.ebi.ac.uk/ols/ontologies/uberon/terms?iri=http%3A%2F%2Fpurl.obolibrary.org%2Fobo%2FUBERON_">liver</sd-tag> lysates analyzed by <sd-tag id="sdTag262" source="sdapi" category="assay" entity_type="" role="" text="immunoblotting" 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&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotting</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=2418"/></sd-panel><sd-panel panel_id="2419">C Quantification of <sd-tag id="sdTag266" source="sdapi" category="entity" entity_type="protein" role="assayed" text="S6" ext_ids="P62754" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="Rps6" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="https://www.uniprot.org/uniprot/">S6</sd-tag> phosphorylation of <sd-tag id="sdTag268" source="sdapi" category="entity" entity_type="tissue" role="component" text="brain" ext_ids="UBERON:0000955" norm_text="" ext_dbs="Uberon" in_caption="True" ext_names="brain" ext_tax_ids="" ext_tax_names="" ext_urls="https://www.ebi.ac.uk/ols/ontologies/uberon/terms?iri=http%3A%2F%2Fpurl.obolibrary.org%2Fobo%2FUBERON_">brain</sd-tag> lysates from WT (n = 2) and <sd-tag id="sdTag269" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Aldh5a1" ext_ids="214579" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Aldh5a1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Aldh5a1</sd-tag>−/− <sd-tag id="sdTag270" source="sdapi" category="entity" entity_type="organism" role="component" text="mice" ext_ids="10090" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Mus musculus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">mice</sd-tag> treated with vehicle (n = 3) or <sd-tag id="sdTag271" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="rapamycin" ext_ids="CHEBI:9168" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="sirolimus" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">rapamycin</sd-tag> (n = 3) after normalization (WT set to 1).<graphic href="https://api.sourcedata.io/file.php?panel_id=2419"/></sd-panel><sd-panel panel_id="2420">D <sd-tag id="sdTag272" source="sdapi" category="entity" entity_type="protein" role="assayed" text="S6" ext_ids="P62754" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="Rps6" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="https://www.uniprot.org/uniprot/">S6</sd-tag> phosphorylation of <sd-tag id="sdTag274" source="sdapi" category="entity" entity_type="tissue" role="component" text="brain" ext_ids="UBERON:0000955" norm_text="" ext_dbs="Uberon" in_caption="True" ext_names="brain" ext_tax_ids="" ext_tax_names="" ext_urls="https://www.ebi.ac.uk/ols/ontologies/uberon/terms?iri=http%3A%2F%2Fpurl.obolibrary.org%2Fobo%2FUBERON_">brain</sd-tag> lysates analyzed by <sd-tag id="sdTag275" source="sdapi" category="assay" entity_type="" role="" text="immunoblotting" 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&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotting</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=2420"/></sd-panel></fig></article>