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<article doi="10.1002/eji.201242835" abstract="Legionella pneumophila (L. pneumophila) is an intracellular bacterium of human alveolar macrophages that causes Legionnaires' disease. In contrast to humans, most inbred mouse strains are restrictive to L. pneumophila replication. We demonstrate that autophagy targets L. pneumophila vacuoles to lysosomes and that this process requires ubiquitination of L. pneumophila vacuoles and the subsequent binding of the autophagic adaptor p62/SQSTM1 to ubiquitinated vacuoles. The L. pneumophila legA9 encodes for an ankyrin-containing protein with unknown role. We show that the legA9 mutant replicate in WT mice and their bone marrow-derived macrophages. This is the first L. pneumophila mutant to be found to replicate in WT bone marrow-derived macrophages other than the Fla mutant. Less legA9 mutant-containing vacuoles acquired ubiquitin labeling and p62/SQSTM1 staining, evading autophagy uptake and avoiding lysosomal fusion. Thus, we describe a bacterial protein that targets the L. pneumophila-containing vacuole for autophagy uptake."><fig id="1091"><title></title><label>Figure 1</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1091"/><sd-panel panel_id="2125"><div><span><strong>(A)</strong> <sd-tag id="sdTag90" source="sdapi" category="entity" entity_type="cell_type" role="component" text="BMDMs" ext_ids="CL:0002476" norm_text="" ext_dbs="CL" in_caption="True" ext_names="bone marrow macrophage" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">BMDMs</sd-tag> were infected with</span> <em><sd-tag id="sdTag92" source="sdapi" category="entity" entity_type="organism" role="assayed" text="L. pneumophila" ext_ids="446" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Legionella pneumophila" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">L. pneumophila</sd-tag></em> <span><sd-tag id="sdTag93" source="sdapi" category="entity" entity_type="gene" role="experiment" text="JR32" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">JR32</sd-tag> or the</span> <em><sd-tag id="sdTag95" source="sdapi" category="entity" entity_type="gene" role="experiment" text="legA9" ext_ids="19831968" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="legA9" ext_tax_ids="272624" ext_tax_names="Legionella pneumophila subsp. pneumophila str. Philadelphia 1" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">legA9</sd-tag></em> <span>mutant with MOI of 0.5. CFUs were scored at 1, 24, 48, and 72 h. <strong>(B)</strong> <sd-tag id="sdTag330" source="sdapi" category="entity" entity_type="cell_type" role="component" text="BMDMs" ext_ids="CL:0002476" norm_text="" ext_dbs="CL" in_caption="True" ext_names="bone marrow macrophage" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">BMDMs</sd-tag> were infected with <sd-tag id="sdTag94" source="sdapi" category="entity" entity_type="gene" role="experiment" text="JR32" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">JR32</sd-tag> or the</span> <em><sd-tag id="sdTag97" source="sdapi" category="entity" entity_type="gene" role="experiment" text="legA9" ext_ids="19831968" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="legA9" ext_tax_ids="272624" ext_tax_names="Legionella pneumophila subsp. pneumophila str. Philadelphia 1" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">legA9</sd-tag></em> <span>mutant harboring empty PL, or</span> <em><sd-tag id="sdTag98" source="sdapi" category="entity" entity_type="gene" role="experiment" text="legA9" ext_ids="19831968" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="legA9" ext_tax_ids="272624" ext_tax_names="Legionella pneumophila subsp. pneumophila str. Philadelphia 1" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">legA9</sd-tag></em><span>+</span> <em><sd-tag id="sdTag99" source="sdapi" category="entity" entity_type="gene" role="experiment" text="legA9" ext_ids="19831968" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="legA9" ext_tax_ids="272624" ext_tax_names="Legionella pneumophila subsp. pneumophila str. Philadelphia 1" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">legA9</sd-tag></em> <span>pBC‐KS</span><span>+</span> <span>PL. (A, B) Data are presented as means ± SD of at least three independent experiments. Asterisks indicate significant differences, (*</span><em>p</em> <span>< 0.05, **</span><em>p</em> <span>< 0.01, ***</span><em>p</em> <span>< 0.001; two‐tailed</span> <em>t</em><span>‐test).</span></div><graphic href="https://api.sourcedata.io/file.php?panel_id=2125"/></sd-panel><sd-panel panel_id="2127"><p>Four female <sd-tag id="sdTag310" source="sdapi" category="entity" entity_type="organism" role="component" text="C57BL/6 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/">C57BL/6 mice</sd-tag>/group received 1 × 10<sup>6</sup> of <sd-tag id="sdTag309" source="sdapi" category="entity" entity_type="gene" role="experiment" text="JR32" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">JR32</sd-tag> or <em><sd-tag id="sdTag312" source="sdapi" category="entity" entity_type="gene" role="experiment" text="legA9" ext_ids="19831968" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="legA9" ext_tax_ids="272624" ext_tax_names="Legionella pneumophila subsp. pneumophila str. Philadelphia 1" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">legA9</sd-tag></em> mutant bacteria intratracheally. <sd-tag id="sdTag311" source="sdapi" category="entity" entity_type="tissue" role="component" text="Lungs" ext_ids="UBERON:0002048" norm_text="" ext_dbs="Uberon" in_caption="True" ext_names="lung" 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_">Lungs</sd-tag> were homogenized and plated for <sd-tag id="sdTag308" source="sdapi" category="assay" entity_type="" role="" text="CFU" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">CFU</sd-tag>s counting at (C) 4 or (D) 48 h postinfection. (C, D) Data are shown as mean + SD of <em>n</em> = 4 and are representative as one of two independent experiments. Asterisks indicate significant differences (**<em>p</em> < 0.01 two‐tailed <em>t</em>‐test)</p><graphic href="https://api.sourcedata.io/file.php?panel_id=2127"/></sd-panel></fig><fig id="1092"><title></title><label>Figure 2</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1092"/><sd-panel panel_id="2128"><b>(A)</b> Representative images from 6 h infected WT <span><sd-tag id="sdTag331" source="sdapi" category="entity" entity_type="cell_type" role="component" text="BMDMs" ext_ids="CL:0002476" norm_text="" ext_dbs="CL" in_caption="True" ext_names="bone marrow macrophage" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">BMDMs</sd-tag></span> with <sd-tag id="sdTag21" source="sdapi" category="entity" entity_type="gene" role="experiment" text="JR32" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">JR32</sd-tag> or <i><sd-tag id="sdTag22" source="sdapi" category="entity" entity_type="gene" role="experiment" text="legA9" ext_ids="19831968" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="legA9" ext_tax_ids="272624" ext_tax_names="Legionella pneumophila subsp. pneumophila str. Philadelphia 1" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">legA9</sd-tag></i> mutant bacteria. <sd-tag id="sdTag29" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="Nuclei" 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/">Nuclei</sd-tag> were stained blue with <sd-tag id="sdTag273" 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> and <i><sd-tag id="sdTag23" source="sdapi" category="entity" entity_type="organism" role="assayed" text="L. pneumophila" ext_ids="446" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Legionella pneumophila" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">L. pneumophila</sd-tag></i> stained green with <i><sd-tag id="sdTag24" source="sdapi" category="entity" entity_type="organism" role="assayed" text="L. pneumophila" ext_ids="446" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Legionella pneumophila" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">L. pneumophila</sd-tag></i> antibody. <sd-tag id="sdTag274" source="sdapi" category="assay" entity_type="" role="" text="Lyso−tracker" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Lyso−tracker</sd-tag> red was used to stain acidified <sd-tag id="sdTag32" 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>. White arrows show <i><sd-tag id="sdTag25" source="sdapi" category="entity" entity_type="organism" role="assayed" text="L. pneumophila" ext_ids="446" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Legionella pneumophila" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">L. pneumophila</sd-tag></i> colocalization with lysotracker.<graphic href="https://api.sourcedata.io/file.php?panel_id=2128"/></sd-panel><sd-panel panel_id="2129"><b>(B)</b> Percent colocalization of <i><sd-tag id="sdTag33" source="sdapi" category="entity" entity_type="organism" role="assayed" text="L. pneumophila" ext_ids="446" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Legionella pneumophila" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">L. pneumophila</sd-tag></i> with <sd-tag id="sdTag275" source="sdapi" category="assay" entity_type="" role="" text="lyso‐tracker" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">lyso‐tracker</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=2129"/></sd-panel><sd-panel panel_id="2130"><b>(C)</b> Representative images from 6 h infected WT <sd-tag id="sdTag46" source="sdapi" category="entity" entity_type="cell_type" role="component" text="BMDMs" ext_ids="CL:0002476" norm_text="" ext_dbs="CL" in_caption="True" ext_names="bone marrow macrophage" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">BMDMs</sd-tag> with <sd-tag id="sdTag43" source="sdapi" category="entity" entity_type="gene" role="experiment" text="JR32" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">JR32</sd-tag> or <i><sd-tag id="sdTag44" source="sdapi" category="entity" entity_type="gene" role="experiment" text="legA9" ext_ids="19831968" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="legA9" ext_tax_ids="272624" ext_tax_names="Legionella pneumophila subsp. pneumophila str. Philadelphia 1" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">legA9</sd-tag></i> mutant bacteria. <sd-tag id="sdTag103" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9CQV6///Q91VR7" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="Map1lc3b///Map1lc3a" ext_tax_ids="10090///10090" ext_tax_names="Mus musculus///Mus musculus" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag> is stained red with <sd-tag id="sdTag104" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9CQV6///Q91VR7" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="Map1lc3b///Map1lc3a" ext_tax_ids="10090///10090" ext_tax_names="Mus musculus///Mus musculus" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag> antibody. White arrows show <sd-tag id="sdTag236" source="sdapi" category="entity" entity_type="organism" role="assayed" text="L. pneumophila" ext_ids="446" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Legionella pneumophila" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/"><i>L. pneumophila</i></sd-tag> colocalization with <sd-tag id="sdTag105" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9CQV6///Q91VR7" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="Map1lc3b///Map1lc3a" ext_tax_ids="10090///10090" ext_tax_names="Mus musculus///Mus musculus" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=2130"/></sd-panel><sd-panel panel_id="2131"><b>(D)</b> Percent colocalization of <i><sd-tag id="sdTag50" source="sdapi" category="entity" entity_type="organism" role="assayed" text="L. pneumophila" ext_ids="446" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Legionella pneumophila" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">L. pneumophila</sd-tag></i> with <sd-tag id="sdTag106" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9CQV6///Q91VR7" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="Map1lc3b///Map1lc3a" ext_tax_ids="10090///10090" ext_tax_names="Mus musculus///Mus musculus" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag>. Hundred bacteria were scored from each coverslips. Images were captured with the 60× objective and magnified 3×, scale bar = 10 μm. (A-D) Data are presented as mean + SD of <i>n</i> = 3 and are representative of three independent experiments. Asterisks indicate significant differences (**<i>p</i> < 0.01, ***<i>p</i> < 0.001; two‐tailed <i>t</i>‐test).<graphic href="https://api.sourcedata.io/file.php?panel_id=2131"/></sd-panel></fig><fig id="1093"><title></title><label>Figure 3</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1093"/><sd-panel panel_id="2132"><p>Induction of autophagy in <sd-pretag id="sdPretag14449969399060" parent-tag-id="339">macrophages</sd-pretag> restricts <em><sd-tag id="sdTag304" source="sdapi" category="entity" entity_type="gene" role="experiment" text="legA9" ext_ids="19831968" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="legA9" ext_tax_ids="272624" ext_tax_names="Legionella pneumophila subsp. pneumophila str. Philadelphia 1" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">legA9</sd-tag></em> mutant replication, and rescues colocalization with <sd-tag id="sdTag302" source="sdapi" category="entity" entity_type="protein" role="component" text="LC3" ext_ids="Q9CQV6///Q91VR7" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="Map1lc3b///Map1lc3a" ext_tax_ids="10090///10090" ext_tax_names="Mus musculus///Mus musculus" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag> and fusion with <sd-tag id="sdTag303" source="sdapi" category="entity" entity_type="subcellular" role="component" 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>. <sd-tag id="sdTag332" source="sdapi" category="entity" entity_type="cell_type" role="component" text="BMDMs" ext_ids="CL:0002476" norm_text="" ext_dbs="CL" in_caption="True" ext_names="bone marrow macrophage" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">BMDMs</sd-tag> were untreated or pretreated with <sd-tag id="sdTag68" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="rapamycin (Rap)" 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 (Rap)</sd-tag> for 1 h and kept throughout the infection. <sd-tag id="sdTag63" source="sdapi" category="entity" entity_type="cell_type" role="component" text="BMDMs" ext_ids="CL:0002476" norm_text="" ext_dbs="CL" in_caption="True" ext_names="bone marrow macrophage" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">BMDMs</sd-tag> were infected with <em><sd-tag id="sdTag64" source="sdapi" category="entity" entity_type="organism" role="assayed" text="L. pneumophila" ext_ids="446" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Legionella pneumophila" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">L. pneumophila</sd-tag></em> <sd-tag id="sdTag65" source="sdapi" category="entity" entity_type="gene" role="experiment" text="JR32" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">JR32</sd-tag> or the <em><sd-tag id="sdTag280" source="sdapi" category="entity" entity_type="gene" role="experiment" text="leg A9" ext_ids="19831968" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="legA9" ext_tax_ids="272624" ext_tax_names="Legionella pneumophila subsp. pneumophila str. Philadelphia 1" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">leg A9</sd-tag></em> mutant for (A) 1 or (B) 24 h with an MOI of 0.5. <sd-tag id="sdTag66" source="sdapi" category="assay" entity_type="" role="" text="CFUs" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">CFUs</sd-tag> were scored at these time points.</p><graphic href="https://api.sourcedata.io/file.php?panel_id=2132"/></sd-panel><sd-panel panel_id="2133"><b>(C, D)</b> Percent colocalization of <i><sd-tag id="sdTag71" source="sdapi" category="entity" entity_type="organism" role="assayed" text="L. pneumophila" ext_ids="446" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Legionella pneumophila" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">L. pneumophila</sd-tag></i> with (C) <sd-tag id="sdTag281" source="sdapi" category="assay" entity_type="" role="" text="lyso‐tracker" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">lyso‐tracker</sd-tag> and (D) <sd-tag id="sdTag73" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9CQV6///Q91VR7" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="Map1lc3b///Map1lc3a" ext_tax_ids="10090///10090" ext_tax_names="Mus musculus///Mus musculus" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag>. Hundred bacteria were scored from each coverslips. (A-D) Data are presented as mean + SD of <i>n</i> = 3 and are representative of three independent experiments. Asterisks indicate significant differences (**<i>p</i> < 0.01; two‐tailed <i>t</i>‐test).<graphic href="https://api.sourcedata.io/file.php?panel_id=2133"/></sd-panel></fig><fig id="1094"><title></title><label>Figure 4</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1094"/><sd-panel panel_id="2134"><b>(A)</b> <span><sd-tag id="sdTag333" source="sdapi" category="entity" entity_type="cell_type" role="component" text="BMDMs" ext_ids="CL:0002476" norm_text="" ext_dbs="CL" in_caption="True" ext_names="bone marrow macrophage" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">BMDMs</sd-tag> derived from WT and</span> <sd-tag id="sdTag88" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Atg5" ext_ids="11793" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Atg5" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Atg5</sd-tag><sup>−/−</sup> <sd-tag id="sdTag84" 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 infected with <i><sd-tag id="sdTag85" source="sdapi" category="entity" entity_type="organism" role="assayed" text="L. pneumophila" ext_ids="446" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Legionella pneumophila" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">L. pneumophila</sd-tag></i> <sd-tag id="sdTag86" source="sdapi" category="entity" entity_type="gene" role="experiment" text="JR32" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">JR32</sd-tag> or <i><sd-tag id="sdTag87" source="sdapi" category="entity" entity_type="gene" role="experiment" text="legA9" ext_ids="19831968" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="legA9" ext_tax_ids="272624" ext_tax_names="Legionella pneumophila subsp. pneumophila str. Philadelphia 1" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">legA9</sd-tag></i> mutant at an MOI of 0.5. Data are shown as mean ± SD of <i>n</i> = 3 and are representative of two independent experiments. Asterisks indicate significant differences (*<i>p</i> <span>< 0.05; two−tailed</span> <i>t</i>−test.<graphic href="https://api.sourcedata.io/file.php?panel_id=2134"/></sd-panel><sd-panel panel_id="2135"><b>(B)</b> <sd-tag id="sdTag109" source="sdapi" category="assay" entity_type="" role="" text="Western blot" 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">Western blot</sd-tag> showing the absence of <sd-tag id="sdTag246" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Atg 5" ext_ids="11793" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Atg5" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Atg 5</sd-tag> in the knockout. Data are representative of three independent experiments.<graphic href="https://api.sourcedata.io/file.php?panel_id=2135"/></sd-panel></fig><fig id="1095"><title></title><label>Figure 5</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1095"/><sd-panel panel_id="2136"><b>(A)</b> Representative images from 6 h infected <span><sd-tag id="sdTag334" source="sdapi" category="entity" entity_type="cell_type" role="component" text="BMDMs" ext_ids="CL:0002476" norm_text="" ext_dbs="CL" in_caption="True" ext_names="bone marrow macrophage" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">BMDMs</sd-tag></span> with <sd-tag id="sdTag114" source="sdapi" category="entity" entity_type="gene" role="experiment" text="JR32" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">JR32</sd-tag> or <i><sd-tag id="sdTag115" source="sdapi" category="entity" entity_type="gene" role="experiment" text="legA9" ext_ids="19831968" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="legA9" ext_tax_ids="272624" ext_tax_names="Legionella pneumophila subsp. pneumophila str. Philadelphia 1" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">legA9</sd-tag></i> mutant bacteria. <sd-tag id="sdTag116" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Polyubiquitin" ext_ids="Q6ZWU9///P62984///P0CG50///P0CG49" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="Rps27///Uba52///Ubc///Ubb" ext_tax_ids="10090///10090///10090///10090" ext_tax_names="Mus musculus///Mus musculus///Mus musculus///Mus musculus" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Polyubiquitin</sd-tag> is stained red. White arrows show <i><sd-tag id="sdTag120" source="sdapi" category="entity" entity_type="organism" role="assayed" text="L. pneumophila" ext_ids="446" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Legionella pneumophila" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">L. pneumophila</sd-tag></i> colocalization with <sd-tag id="sdTag318" source="sdapi" category="entity" entity_type="protein" role="assayed" text="polyubiquitin" ext_ids="P0CG50///P0CG49///Q6ZWU9///P62984" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="Ubc///Ubb///Rps27///Uba52" ext_tax_ids="10090///10090///10090///10090" ext_tax_names="Mus musculus///Mus musculus///Mus musculus///Mus musculus" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">polyubiquitin</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=2136"/></sd-panel><sd-panel panel_id="2137"><b>(B)</b> Percent colocalization of <i><sd-tag id="sdTag121" source="sdapi" category="entity" entity_type="organism" role="assayed" text="L. pneumophila" ext_ids="446" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Legionella pneumophila" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">L. pneumophila</sd-tag></i> with <sd-tag id="sdTag128" source="sdapi" category="entity" entity_type="protein" role="assayed" text="polyubiquitin" ext_ids="Q6ZWU9///P62984///P0CG50///P0CG49" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="Rps27///Uba52///Ubc///Ubb" ext_tax_ids="10090///10090///10090///10090" ext_tax_names="Mus musculus///Mus musculus///Mus musculus///Mus musculus" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">polyubiquitin</sd-tag>. Hundred bacteria were scored from each coverslips. Images were captured with the 60× objective and magnified 3×, scale bar = 10 μm. Data are presented as means + SD of <i>n</i> = 3 and are representative of two independent experiments. Asterisks indicate significant differences (*<i>p</i> < 0.05, **<i>p</i> < 0.01, ***<i>p</i> < 0.01; two‐tailed <i>t</i>‐test).<graphic href="https://api.sourcedata.io/file.php?panel_id=2137"/></sd-panel></fig><fig id="1096"><title></title><label>Figure 6</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1096"/><sd-panel panel_id="2138"><b>(A)</b> Representative images from 6 h <span>infected <sd-tag id="sdTag335" source="sdapi" category="entity" entity_type="cell_type" role="component" text="BMDMs" ext_ids="CL:0002476" norm_text="" ext_dbs="CL" in_caption="True" ext_names="bone marrow macrophage" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">BMDMs</sd-tag> with</span> <sd-tag id="sdTag131" source="sdapi" category="entity" entity_type="gene" role="experiment" text="JR32" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">JR32</sd-tag> or <i><sd-tag id="sdTag132" source="sdapi" category="entity" entity_type="gene" role="experiment" text="legA9" ext_ids="19831968" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="legA9" ext_tax_ids="272624" ext_tax_names="Legionella pneumophila subsp. pneumophila str. Philadelphia 1" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">legA9</sd-tag></i> mutant bacteria. <sd-tag id="sdTag133" source="sdapi" category="entity" entity_type="protein" role="assayed" text="P62" ext_ids="Q64337" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="Sqstm1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="https://www.uniprot.org/uniprot/">P62</sd-tag> is stained red with <sd-tag id="sdTag139" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p62" ext_ids="Q64337" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="Sqstm1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="https://www.uniprot.org/uniprot/">p62</sd-tag> antibody. White arrows show <i><sd-tag id="sdTag134" source="sdapi" category="entity" entity_type="organism" role="assayed" text="L. pneumophila" ext_ids="446" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Legionella pneumophila" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">L. pneumophila</sd-tag></i> colocalization with <sd-tag id="sdTag138" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p62" ext_ids="Q64337" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="Sqstm1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="https://www.uniprot.org/uniprot/">p62</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=2138"/></sd-panel><sd-panel panel_id="2139"><b>(B)</b> Percent colocalization of <sd-tag id="sdTag150" source="sdapi" category="entity" entity_type="organism" role="assayed" text="L. pneumophila" ext_ids="446" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Legionella pneumophila" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/"><i>L. pneumophila</i></sd-tag> with <sd-tag id="sdTag141" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p62" ext_ids="Q64337" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="Sqstm1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="https://www.uniprot.org/uniprot/">p62</sd-tag>. Hundred bacteria were scored from each coverslips. Images were captured with the 60× objective and magnified 3×, scale bar = 10 μm.<graphic href="https://api.sourcedata.io/file.php?panel_id=2139"/></sd-panel><sd-panel panel_id="2140"><p><strong>(C)</strong><sd-tag id="sdTag151" source="sdapi" category="entity" entity_type="cell_type" role="component" text="BMDMs" ext_ids="CL:0000235///CL:0002476" norm_text="" ext_dbs="CL///CL" in_caption="True" ext_names="macrophage///bone marrow macrophage" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org//// https://identifiers.org/">BMDMs</sd-tag> were infected with <sd-tag id="sdTag152" source="sdapi" category="entity" entity_type="gene" role="experiment" text="JR32" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">JR32</sd-tag> or the <em><sd-tag id="sdTag153" source="sdapi" category="entity" entity_type="gene" role="experiment" text="legA9" ext_ids="19831968" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="legA9" ext_tax_ids="272624" ext_tax_names="Legionella pneumophila subsp. pneumophila str. Philadelphia 1" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">legA9</sd-tag></em> mutant for 6 h and the level of <sd-tag id="sdTag154" source="sdapi" category="entity" entity_type="protein" role="assayed" text="p62" ext_ids="Q64337" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="Sqstm1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="https://www.uniprot.org/uniprot/">p62</sd-tag> protein was quantified in cell lysates by <sd-tag id="sdTag157" source="sdapi" category="assay" entity_type="" role="" text="ELISA" ext_ids="BAO_0000134" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="ELISA" 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">ELISA</sd-tag> (performed in triplicate and the SD are too small to detect).</p><graphic href="https://api.sourcedata.io/file.php?panel_id=2140"/></sd-panel><sd-panel panel_id="2141"><p><span><sd-tag id="sdTag158" source="sdapi" category="entity" entity_type="cell_type" role="component" text="BMDMs" ext_ids="CL:0000235///CL:0002476" norm_text="" ext_dbs="CL///CL" in_caption="True" ext_names="macrophage///bone marrow macrophage" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org//// https://identifiers.org/">BMDMs</sd-tag> were nucleofected with scrambled control (sictrl) or siRNA against <sd-tag id="sdTag159" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p62" ext_ids="18412" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Sqstm1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p62</sd-tag> (sip 62). Twenty‐four hours after nucleofection, murine <sd-tag id="sdTag340" source="sdapi" category="entity" entity_type="cell_type" role="component" text="macrophages" ext_ids="CL:0000235" norm_text="" ext_dbs="CL" in_caption="True" ext_names="macrophage" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">macrophages</sd-tag> were infected with <sd-tag id="sdTag160" source="sdapi" category="entity" entity_type="gene" role="experiment" text="JR32" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">JR32</sd-tag> or</span> <em><sd-tag id="sdTag161" source="sdapi" category="entity" entity_type="gene" role="experiment" text="legA9" ext_ids="19831968" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="legA9" ext_tax_ids="272624" ext_tax_names="Legionella pneumophila subsp. pneumophila str. Philadelphia 1" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">legA9</sd-tag></em> <span>mutant with an MOI of 0.5 for 1 h (D and F) or 24 h (E and G).</span></p><graphic href="https://api.sourcedata.io/file.php?panel_id=2141"/></sd-panel><sd-panel panel_id="2142"><b>(H)</b> <sd-tag id="sdTag166" source="sdapi" category="assay" entity_type="" role="" text="Western blot" 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">Western blot</sd-tag> analysis showing the downregulation of <sd-tag id="sdTag167" source="sdapi" category="entity" entity_type="gene" role="intervention" text="p62" ext_ids="18412" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Sqstm1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">p62</sd-tag> at 24 and 48 h postnucleofection. (A-B and D-H) Data are shown as mean + SD of <i>n</i> = 3 and are representative of three independent experiments. (C) Data are representative of one experiment <i>n</i> = 3. Asterisks indicate significant differences (*<i>p</i> <span>< 0.05, **</span><i>p</i> <span>< 0.01; two−tailed</span> <i>t</i>−test).<graphic href="https://api.sourcedata.io/file.php?panel_id=2142"/></sd-panel></fig><fig id="1097"><title></title><label>Figure 7</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1097"/><sd-panel panel_id="2143"><b>(A)</b> <sd-tag id="sdTag175" source="sdapi" category="entity" entity_type="gene" role="component" text="Caspase−1" ext_ids="12362" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Casp1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Caspase−1</sd-tag> KO (casp−1<sup>−/−</sup>) <span><sd-tag id="sdTag339" source="sdapi" category="entity" entity_type="cell_type" role="component" text="macrophages" ext_ids="CL:0000235" norm_text="" ext_dbs="CL" in_caption="True" ext_names="macrophage" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">macrophages</sd-tag></span> were infected with <i><sd-tag id="sdTag172" source="sdapi" category="entity" entity_type="organism" role="assayed" text="L. pneumophila" ext_ids="446" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Legionella pneumophila" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">L. pneumophila</sd-tag></i> <sd-tag id="sdTag173" source="sdapi" category="entity" entity_type="gene" role="experiment" text="JR32" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">JR32</sd-tag> or <i><sd-tag id="sdTag174" source="sdapi" category="entity" entity_type="gene" role="experiment" text="legA9" ext_ids="19831968" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="legA9" ext_tax_ids="272624" ext_tax_names="Legionella pneumophila subsp. pneumophila str. Philadelphia 1" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">legA9</sd-tag></i> with an MOI of 0.5 for 1, 24, 48, and 72 h.<graphic href="https://api.sourcedata.io/file.php?panel_id=2143"/></sd-panel><sd-panel panel_id="2144"><p><strong>(B)</strong> <sd-tag id="sdTag182" source="sdapi" category="assay" entity_type="" role="" text="CFUs" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">CFUs</sd-tag> were measured at the indicated time points. Levels of <sd-tag id="sdTag266" source="sdapi" category="entity" entity_type="protein" role="assayed" text="IL‐1β" ext_ids="P10749" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="Il1b" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="https://www.uniprot.org/uniprot/">IL‐1β</sd-tag> were detected in supernatants of WT or <sd-tag id="sdTag189" source="sdapi" category="entity" entity_type="gene" role="intervention" text="casp‐1" ext_ids="12362" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="Casp1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">casp‐1</sd-tag><sup>−/−</sup> <sd-tag id="sdTag336" source="sdapi" category="entity" entity_type="cell_type" role="component" text="BMDMs" ext_ids="CL:0002476" norm_text="" ext_dbs="CL" in_caption="True" ext_names="bone marrow macrophage" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">BMDMs</sd-tag> infected with <sd-tag id="sdTag192" source="sdapi" category="entity" entity_type="gene" role="experiment" text="JR32" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">JR32</sd-tag> or the <em><sd-tag id="sdTag194" source="sdapi" category="entity" entity_type="gene" role="experiment" text="legA9" ext_ids="19831968" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="legA9" ext_tax_ids="272624" ext_tax_names="Legionella pneumophila subsp. pneumophila str. Philadelphia 1" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">legA9</sd-tag></em> mutant after 24 h. WT <sd-tag id="sdTag185" source="sdapi" category="entity" entity_type="cell_type" role="component" text="BMDMs" ext_ids="CL:0000235///CL:0002476" norm_text="" ext_dbs="CL///CL" in_caption="True" ext_names="macrophage///bone marrow macrophage" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org//// https://identifiers.org/">BMDMs</sd-tag> were either not treated (NT) or infected with <em><sd-tag id="sdTag186" source="sdapi" category="entity" entity_type="organism" role="intervention" text="L. pneumophila" ext_ids="446" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Legionella pneumophila" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">L. pneumophila</sd-tag></em> <sd-tag id="sdTag187" source="sdapi" category="entity" entity_type="gene" role="experiment" text="JR32" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">JR32</sd-tag> or <em><sd-tag id="sdTag193" source="sdapi" category="entity" entity_type="gene" role="experiment" text="legA9" ext_ids="19831968" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="legA9" ext_tax_ids="272624" ext_tax_names="Legionella pneumophila subsp. pneumophila str. Philadelphia 1" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">legA9</sd-tag></em> mutant bacteria for 2 h.</p><graphic href="https://api.sourcedata.io/file.php?panel_id=2144"/></sd-panel><sd-panel panel_id="2145"><p>Activation of <sd-tag id="sdTag196" source="sdapi" category="entity" entity_type="protein" role="assayed" text="casp‐1" ext_ids="P29452" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="Casp1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="https://www.uniprot.org/uniprot/">casp‐1</sd-tag> or <sd-tag id="sdTag199" source="sdapi" category="entity" entity_type="protein" role="assayed" text="casp‐7" ext_ids="P97864" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="Casp7" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="https://www.uniprot.org/uniprot/">casp‐7</sd-tag> was detected in cell extracts using (C) <sd-tag id="sdTag197" source="sdapi" category="entity" entity_type="protein" role="assayed" text="casp‐1" ext_ids="P29452" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="Casp1" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="https://www.uniprot.org/uniprot/">casp‐1</sd-tag> or (D) <sd-tag id="sdTag198" source="sdapi" category="entity" entity_type="protein" role="assayed" text="casp‐7" ext_ids="P97864" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="Casp7" ext_tax_ids="10090" ext_tax_names="Mus musculus" ext_urls="https://www.uniprot.org/uniprot/">casp‐7</sd-tag> antibodies, respectively. (A-D) Data are shown as mean ± SD of <i>n</i> = 3 and are representative of three independent experiments.</p><graphic href="https://api.sourcedata.io/file.php?panel_id=2145"/></sd-panel></fig><fig id="1098"><title></title><label>Figure 8</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1098"/><sd-panel panel_id="2146"><b>(A)</b> <span><sd-tag id="sdTag337" source="sdapi" category="entity" entity_type="cell_type" role="assayed" text="BMDMs" ext_ids="CL:0002476" norm_text="" ext_dbs="CL" in_caption="True" ext_names="bone marrow macrophage" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">BMDMs</sd-tag></span> were not infected (NT) or infected with <i><sd-tag id="sdTag206" source="sdapi" category="entity" entity_type="organism" role="intervention" text="L. pneumophila" ext_ids="446" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Legionella pneumophila" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">L. pneumophila</sd-tag></i> <sd-tag id="sdTag207" source="sdapi" category="entity" entity_type="gene" role="experiment" text="JR32" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">JR32</sd-tag> or the <i><sd-tag id="sdTag208" source="sdapi" category="entity" entity_type="gene" role="experiment" text="legA9" ext_ids="19831968" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="legA9" ext_tax_ids="272624" ext_tax_names="Legionella pneumophila subsp. pneumophila str. Philadelphia 1" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">legA9</sd-tag></i> mutant at an MOI of 0.5, 1 or 5 for 24 h. Apoptosis was quantified by Photometric enzyme immunoassay analysis of cytoplasmic (apoptosis) histone−associated−DNA fragments (<sd-tag id="sdTag209" source="sdapi" category="assay" entity_type="" role="" text="ELISA" ext_ids="BAO_0000134" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="ELISA" 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">ELISA</sd-tag>).<graphic href="https://api.sourcedata.io/file.php?panel_id=2146"/></sd-panel><sd-panel panel_id="2147"><b>(B)</b> <span><sd-tag id="sdTag338" source="sdapi" category="entity" entity_type="cell_type" role="component" text="BMDMs" ext_ids="CL:0002476" norm_text="" ext_dbs="CL" in_caption="True" ext_names="bone marrow macrophage" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">BMDMs</sd-tag></span> were either not treated (NT) or treated with <i><sd-tag id="sdTag213" source="sdapi" category="entity" entity_type="organism" role="intervention" text="L. pneumophila" ext_ids="446" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="Legionella pneumophila" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">L. pneumophila</sd-tag></i> <sd-tag id="sdTag214" source="sdapi" category="entity" entity_type="gene" role="experiment" text="JR32" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">JR32</sd-tag> or the <i><sd-tag id="sdTag215" source="sdapi" category="entity" entity_type="gene" role="experiment" text="legA9" ext_ids="19831968" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="legA9" ext_tax_ids="272624" ext_tax_names="Legionella pneumophila subsp. pneumophila str. Philadelphia 1" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">legA9</sd-tag></i> mutant at an MOI of 0.5 or 5 for 24 h. The fold change in <sd-tag id="sdTag217" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LDH" ext_ids="P00342///P16125///P06151" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="Ldhc///Ldhb///Ldha" ext_tax_ids="10090///10090///10090" ext_tax_names="Mus musculus///Mus musculus///Mus musculus" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LDH</sd-tag> release was measured from the overall population of <span><sd-tag id="sdTag341" source="sdapi" category="entity" entity_type="cell_type" role="component" text="macrophages" ext_ids="CL:0000235" norm_text="" ext_dbs="CL" in_caption="True" ext_names="macrophage" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">macrophages</sd-tag></span>. (A-B) Data are shown as mean ± SD of <i>n</i> = 3 and are representative of three independent experiments.<graphic href="https://api.sourcedata.io/file.php?panel_id=2147"/></sd-panel></fig></article> |
