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<article doi="10.1002/embr.201337995" abstract="The RabGAP protein TBC1D5 controls cellular endomembrane trafficking processes and binds the retromer subunit VPS29 and the ubiquitin-like protein ATG8 (LC3). Here, we describe that TBC1D5 also associates with ATG9 and the active ULK1 complex during autophagy. Moreover, ATG9 and TBC1D5 interact with clathrin and the AP2 complex. Depletion of TBC1D5 leads to missorting of ATG9 to late endosomes upon activation of autophagy, whereas inhibition of clathrin-mediated endocytosis or AP2 depletion alters ATG9 trafficking and its association with TBC1D5. Taken together, our data show that TBC1D5 and the AP2 complex are important novel regulators of the rerouting of ATG9-containing vesicular carriers toward sites of autophagosome formation."><fig id="744"><title></title><label>Figure 1</label><graphic href="https://api.sourcedata.io/file.php?figure_id=744"/><sd-panel panel_id="1075">A Co‐localization of endogenous <sd-tag id="sdTag4" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TBC1D5" ext_ids="Q92609" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TBC1D5</sd-tag> and <sd-tag id="sdTag5" source="sdapi" category="entity" entity_type="protein" role="assayed" text="VPS35" ext_ids="Q96QK1" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="VPS35" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">VPS35</sd-tag> with <sd-tag id="sdTag6" source="sdapi" category="entity" entity_type="protein" role="reporter" text="mCherry" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">mCherry</sd-tag>‐<sd-tag id="sdTag3" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ATG9A" ext_ids="Q7Z3C6" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG9A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ATG9A</sd-tag>, in steady state and autophagy induced by <sd-tag id="sdTag1" source="sdapi" category="entity" entity_type="subcellular" role="component" text="mTOR" ext_ids="GO:0031932///GO:0031931" norm_text="" ext_dbs="Gene Ontology///Gene Ontology" in_caption="True" ext_names="TORC2 complex///TORC1 complex" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term////http://amigo.geneontology.org/amigo/term/">mTOR</sd-tag> inhibitor <sd-tag id="sdTag2" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="KU0063794" ext_ids="CHEBI:85572" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Ku-0063794" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">KU0063794</sd-tag> (6 h).<graphic href="https://api.sourcedata.io/file.php?panel_id=1075"/></sd-panel><sd-panel panel_id="1076">B, C Quantification of co‐localization experiments presented in (A).<graphic href="https://api.sourcedata.io/file.php?panel_id=1076"/></sd-panel><sd-panel panel_id="1077">D <sd-tag id="sdTag14" source="sdapi" category="assay" entity_type="" role="" text="Co‐immunoprecipitation" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Co‐immunoprecipitation</sd-tag> of <sd-tag id="sdTag271" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>‐<sd-tag id="sdTag272" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag>‐<sd-tag id="sdTag10" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TBC1D5" ext_ids="Q92609" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TBC1D5</sd-tag> with <sd-tag id="sdTag9" source="sdapi" category="entity" entity_type="protein" role="reporter" text="myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">myc</sd-tag>‐<sd-tag id="sdTag11" source="sdapi" category="entity" entity_type="protein" role="assayed" text="VPS29" ext_ids="Q9UBQ0" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="VPS29" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">VPS29</sd-tag>, transiently overexpressed in <sd-tag id="sdTag12" source="sdapi" category="entity" entity_type="cell_line" role="component" text="293T cells" ext_ids="CVCL_0063" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293T" ext_tax_ids="" ext_tax_names="" ext_urls="https://identifiers.org/cellosaurus:">293T cells</sd-tag>, <sd-tag id="sdTag275" source="sdapi" category="entity" entity_type="molecule" role="component" text="DMSO" ext_ids="CHEBI:28262" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="dimethyl sulfoxide" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DMSO</sd-tag> or <sd-tag id="sdTag13" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="KU0063794" ext_ids="CHEBI:85572" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Ku-0063794" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">KU0063794</sd-tag> (6 h) treated.<graphic href="https://api.sourcedata.io/file.php?panel_id=1077"/></sd-panel><sd-panel panel_id="1078">E, F <sd-tag id="sdTag29" source="sdapi" category="assay" entity_type="" role="" text="Co‐immunoprecipitation" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Co‐immunoprecipitation</sd-tag> of endogenous <sd-tag id="sdTag19" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TBC1D5" ext_ids="Q92609" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TBC1D5</sd-tag> and <sd-tag id="sdTag20" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ATG9A" ext_ids="Q7Z3C6" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG9A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ATG9A</sd-tag> with <sd-tag id="sdTag21" source="sdapi" category="entity" entity_type="protein" role="reporter" text="myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">myc</sd-tag>‐<sd-tag id="sdTag22" source="sdapi" category="entity" entity_type="gene" role="intervention" text="VPS29" ext_ids="51699" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="VPS29" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">VPS29</sd-tag>(E) transiently overexpressed in <sd-tag id="sdTag23" source="sdapi" category="entity" entity_type="cell_line" role="component" text="293T cells" ext_ids="CVCL_0063" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293T" ext_tax_ids="" ext_tax_names="" ext_urls="https://identifiers.org/cellosaurus:">293T cells</sd-tag> or with <sd-tag id="sdTag277" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>‐<sd-tag id="sdTag278" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag>‐<sd-tag id="sdTag226" source="sdapi" category="entity" entity_type="protein" role="assayed" text="VPS35" ext_ids="Q96QK1" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="VPS35" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">VPS35</sd-tag> (F) transiently overexpressed in <sd-tag id="sdTag26" source="sdapi" category="entity" entity_type="cell_line" role="component" text="293T cells" ext_ids="CVCL_0063" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293T" ext_tax_ids="" ext_tax_names="" ext_urls="https://identifiers.org/cellosaurus:">293T cells</sd-tag>, treated with <sd-tag id="sdTag279" source="sdapi" category="entity" entity_type="molecule" role="component" text="DMSO" ext_ids="CHEBI:28262" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="dimethyl sulfoxide" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DMSO</sd-tag> or <sd-tag id="sdTag27" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="KU0063794" ext_ids="CHEBI:85572" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Ku-0063794" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">KU0063794</sd-tag> (6 h).<graphic href="https://api.sourcedata.io/file.php?panel_id=1078"/></sd-panel><sd-panel panel_id="1079">G <sd-tag id="sdTag37" source="sdapi" category="assay" entity_type="" role="" text="Co‐immunoprecipitation" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Co‐immunoprecipitation</sd-tag> of endogenous <sd-tag id="sdTag30" source="sdapi" category="entity" entity_type="protein" role="assayed" text="VPS29" ext_ids="Q9UBQ0" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="VPS29" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">VPS29</sd-tag> and <sd-tag id="sdTag31" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ATG9A" ext_ids="Q7Z3C6" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG9A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ATG9A</sd-tag> with <sd-tag id="sdTag281" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>‐<sd-tag id="sdTag282" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag>‐<sd-tag id="sdTag33" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TBC1D5" ext_ids="Q92609" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TBC1D5</sd-tag>stably expressed in <sd-tag id="sdTag34" source="sdapi" category="entity" entity_type="cell_line" role="component" text="293T cells" ext_ids="CVCL_0063" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293T" ext_tax_ids="" ext_tax_names="" ext_urls="https://identifiers.org/cellosaurus:">293T cells</sd-tag>, treated with <sd-tag id="sdTag283" source="sdapi" category="entity" entity_type="molecule" role="component" text="DMSO" ext_ids="CHEBI:28262" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="dimethyl sulfoxide" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DMSO</sd-tag> or <sd-tag id="sdTag35" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="KU0063794" ext_ids="CHEBI:85572" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Ku-0063794" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">KU0063794</sd-tag> (6 h).<graphic href="https://api.sourcedata.io/file.php?panel_id=1079"/></sd-panel><sd-panel panel_id="1080">H <sd-tag id="sdTag45" source="sdapi" category="assay" entity_type="" role="" text="Co‐immunoprecipitation" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Co‐immunoprecipitation</sd-tag> of endogenous <sd-tag id="sdTag47" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ULK1" ext_ids="O75385" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ULK1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ULK1</sd-tag> and p<sd-tag id="sdTag48" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ATG13" ext_ids="O75143" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG13" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ATG13</sd-tag> (Ser318) with <sd-tag id="sdTag285" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>‐<sd-tag id="sdTag286" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag>‐<sd-tag id="sdTag39" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TBC1D5" ext_ids="Q92609" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TBC1D5</sd-tag>, stably expressed in <sd-tag id="sdTag46" source="sdapi" category="entity" entity_type="cell_line" role="component" text="T‐REx 293T" ext_ids="CVCL_D585" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="T-REx-293" ext_tax_ids="" ext_tax_names="" ext_urls="https://identifiers.org/cellosaurus:">T‐REx 293T</sd-tag> cells, induced by <sd-tag id="sdTag41" source="sdapi" category="entity" entity_type="molecule" role="component" text="doxycycline" ext_ids="CHEBI:50845" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="doxycycline" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">doxycycline</sd-tag> (50 ng/ml) for 20 h prior to <sd-tag id="sdTag287" source="sdapi" category="entity" entity_type="molecule" role="component" text="DMSO" ext_ids="CHEBI:28262" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="dimethyl sulfoxide" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DMSO</sd-tag> or <sd-tag id="sdTag43" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="KU0063794" ext_ids="CHEBI:85572" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Ku-0063794" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">KU0063794</sd-tag> (6 h) treatment.<graphic href="https://api.sourcedata.io/file.php?panel_id=1080"/></sd-panel><sd-panel panel_id="1081">I Magnified still images extracted from Supplementary Movie S2. <sd-tag id="sdTag49" source="sdapi" category="entity" entity_type="molecule" role="reporter" text="mCherry" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">mCherry</sd-tag>‐<sd-tag id="sdTag50" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TBC1D5" ext_ids="Q92609" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TBC1D5</sd-tag> co‐localizes with <sd-tag id="sdTag51" source="sdapi" category="entity" entity_type="gene" 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="sdTag52" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ATG9" ext_ids="Q7Z3C6" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG9A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ATG9</sd-tag> upon autophagy induction by <sd-tag id="sdTag290" source="sdapi" category="entity" entity_type="subcellular" role="component" text="mTOR" ext_ids="GO:0031932///GO:0031931" norm_text="" ext_dbs="Gene Ontology///Gene Ontology" in_caption="True" ext_names="TORC2 complex///TORC1 complex" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term////http://amigo.geneontology.org/amigo/term/">mTOR</sd-tag> inhibitor, <sd-tag id="sdTag53" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="KU0063794" ext_ids="CHEBI:85572" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Ku-0063794" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">KU0063794</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=1081"/></sd-panel><sd-panel panel_id="15074">H <sd-tag id="sdTag392" source="sdapi" category="assay" entity_type="" role="" text="Co-immunoprecipitation" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Co-immunoprecipitation</sd-tag> of endogenous <sd-tag id="sdTag393" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ULK1" ext_ids="O75385" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ULK1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ULK1</sd-tag> and p<sd-tag id="sdTag391" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ATG13" ext_ids="O75143" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG13" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ATG13</sd-tag> (Ser318) with <sd-tag id="sdTag394" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag395" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag>-<sd-tag id="sdTag396" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TBC1D5" ext_ids="Q92609" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TBC1D5</sd-tag>, stably expressed in <sd-tag id="sdTag390" source="sdapi" category="entity" entity_type="cell_line" role="component" text="T-REx 293T" ext_ids="CVCL_D585" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="T-REx-293" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">T-REx 293T</sd-tag> cells, induced by <sd-tag id="sdTag397" source="sdapi" category="entity" entity_type="molecule" role="component" text="doxycycline" ext_ids="CHEBI:50845" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="doxycycline" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">doxycycline</sd-tag> (50 ng/ml) for 20 h prior to <sd-tag id="sdTag398" source="sdapi" category="entity" entity_type="molecule" role="component" text="DMSO" ext_ids="CHEBI:28262" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="dimethyl sulfoxide" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DMSO</sd-tag> or <sd-tag id="sdTag399" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="KU0063794" ext_ids="CHEBI:85572" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Ku-0063794" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">KU0063794</sd-tag> (6 h) treatment.<graphic href="https://api.sourcedata.io/file.php?panel_id=15074"/></sd-panel></fig><fig id="745"><title></title><label>Figure 2</label><graphic href="https://api.sourcedata.io/file.php?figure_id=745"/><sd-panel panel_id="1082">A Lysates of <sd-tag id="sdTag59" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="" ext_tax_names="" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag> cells stably expressing control shRNA or shRNA targeting <sd-tag id="sdTag60" source="sdapi" category="entity" entity_type="gene" role="intervention" text="VPS29" ext_ids="51699" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="VPS29" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">VPS29</sd-tag> or <sd-tag id="sdTag61" source="sdapi" category="entity" entity_type="gene" role="intervention" text="TBC1D5" ext_ids="9779" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">TBC1D5</sd-tag>. Cells were treated with <sd-tag id="sdTag294" source="sdapi" category="entity" entity_type="molecule" role="component" text="DMSO" ext_ids="CHEBI:28262" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="dimethyl sulfoxide" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DMSO</sd-tag> or <sd-tag id="sdTag62" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="KU0063794" ext_ids="CHEBI:85572" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Ku-0063794" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">KU0063794</sd-tag> (6 h), lysed in RIPA buffer (1% SDS) and lysates were subjected to <sd-tag id="sdTag64" source="sdapi" category="assay" entity_type="" role="" text="SDS–PAGE" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SDS-PAGE</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=1082"/></sd-panel><sd-panel panel_id="1083">B <sd-tag id="sdTag73" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="" ext_tax_names="" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag> cells stably expressing <sd-tag id="sdTag74" source="sdapi" category="entity" entity_type="molecule" role="reporter" text="mCherry" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">mCherry</sd-tag>‐<sd-tag id="sdTag75" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ATG9" ext_ids="Q7Z3C6" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG9A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ATG9</sd-tag> were depleted of <sd-tag id="sdTag76" source="sdapi" category="entity" entity_type="gene" role="intervention" text="VPS29" ext_ids="51699" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="VPS29" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">VPS29</sd-tag> and <sd-tag id="sdTag77" source="sdapi" category="entity" entity_type="gene" role="intervention" text="TBC1D5" ext_ids="9779" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">TBC1D5</sd-tag> using shRNAs, treated with <sd-tag id="sdTag295" source="sdapi" category="entity" entity_type="molecule" role="component" text="DMSO" ext_ids="CHEBI:28262" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="dimethyl sulfoxide" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DMSO</sd-tag> or <sd-tag id="sdTag78" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="KU0063794" ext_ids="CHEBI:85572" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Ku-0063794" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">KU0063794</sd-tag> (6 h), fixed with 2% PFA, and <sd-tag id="sdTag118" source="sdapi" category="assay" entity_type="" role="" text="immunostained" ext_ids="BAO_0002426" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunofluorescent labeling" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunostained</sd-tag> with anti‐<sd-tag id="sdTag79" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LAMP1" ext_ids="P11279" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="LAMP1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">LAMP1</sd-tag> antibody.<graphic href="https://api.sourcedata.io/file.php?panel_id=1083"/></sd-panel><sd-panel panel_id="1084">C Quantification of <sd-tag id="sdTag80" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ATG9" ext_ids="Q7Z3C6" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG9A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ATG9</sd-tag> vesicular structures in cells from (B) under non‐stimulated conditions.<graphic href="https://api.sourcedata.io/file.php?panel_id=1084"/></sd-panel><sd-panel panel_id="1085">D Quantification of <sd-tag id="sdTag83" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ATG9" ext_ids="Q7Z3C6" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG9A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ATG9</sd-tag> and <sd-tag id="sdTag84" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LAMP1" ext_ids="P11279" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="LAMP1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">LAMP1</sd-tag> co‐localization in cells presented in (B).<graphic href="https://api.sourcedata.io/file.php?panel_id=1085"/></sd-panel><sd-panel panel_id="1086">E Quantification of <sd-tag id="sdTag85" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ATG9" ext_ids="Q7Z3C6" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG9A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ATG9</sd-tag> and <sd-tag id="sdTag86" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LAMP1" ext_ids="P11279" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="LAMP1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">LAMP1</sd-tag> co−localization in cells stably depleted for <sd-tag id="sdTag236" source="sdapi" category="entity" entity_type="gene" role="intervention" text="TBC1D5" ext_ids="9779" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">TBC1D5</sd-tag> (shRNA#1) or VPS29 (shRNA#2), transiently transfected with <sd-tag id="sdTag91" source="sdapi" category="entity" entity_type="protein" role="reporter" text="myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">myc</sd-tag>−<sd-tag id="sdTag89" source="sdapi" category="entity" entity_type="gene" role="intervention" text="TBC1D5" ext_ids="9779" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">TBC1D5</sd-tag> plasmid resistant to <sd-pretag id="sdPretag14393827884870" parent-tag-id="95">TBC1D5</sd-pretag> shRNA#1. Cells were treated with <sd-tag id="sdTag90" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="KU0063794" ext_ids="CHEBI:85572" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Ku-0063794" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">KU0063794</sd-tag> (6 h), 20 h post−transfection, fixed, and <sd-tag id="sdTag302" source="sdapi" category="assay" entity_type="" role="" text="immunostained" ext_ids="BAO_0002426" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunofluorescent labeling" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunostained</sd-tag> with anti−LAMP1 and anti−myc antibodies.<graphic href="https://api.sourcedata.io/file.php?panel_id=1086"/></sd-panel><sd-panel panel_id="1087">F Control shRNA and <sd-tag id="sdTag92" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="" ext_tax_names="" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag> cell lysates depleted for <sd-tag id="sdTag93" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TBC1D5" ext_ids="Q92609" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TBC1D5</sd-tag> (shRNA#1), transfected with empty plasmid or with shRNA‐resistant <sd-tag id="sdTag94" source="sdapi" category="entity" entity_type="protein" role="reporter" text="myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">myc</sd-tag>‐<sd-tag id="sdTag95" source="sdapi" category="entity" entity_type="gene" role="intervention" text="TBC1D5" ext_ids="9779" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">TBC1D5</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=1087"/></sd-panel><sd-panel panel_id="1088">G <sd-tag id="sdTag115" source="sdapi" category="assay" entity_type="" role="" text="Immunofluorescence" 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</sd-tag> of <sd-tag id="sdTag98" source="sdapi" category="entity" entity_type="gene" role="component" text="TBC1D5" ext_ids="9779" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">TBC1D5</sd-tag> (shRNA#1) cells transiently transfected with shRNA‐resistant <sd-tag id="sdTag306" source="sdapi" category="entity" entity_type="protein" role="reporter" text="myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">myc</sd-tag>‐<sd-tag id="sdTag99" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TBC1D5" ext_ids="Q92609" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TBC1D5</sd-tag>, treated with <sd-tag id="sdTag100" source="sdapi" category="entity" entity_type="molecule" role="component" text="KU0063794" ext_ids="CHEBI:85572" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Ku-0063794" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">KU0063794</sd-tag> (6 h). 20 h post‐transfection cells were treated, subsequently fixed, and <sd-tag id="sdTag307" source="sdapi" category="assay" entity_type="" role="" text="immunostained" ext_ids="BAO_0002426" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunofluorescent labeling" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunostained</sd-tag> with anti‐<sd-tag id="sdTag102" source="sdapi" category="entity" entity_type="protein" role="reporter" text="myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">myc</sd-tag> and anti‐<sd-tag id="sdTag101" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LAMP1" ext_ids="P11279" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="LAMP1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">LAMP1</sd-tag> antibodies.<graphic href="https://api.sourcedata.io/file.php?panel_id=1088"/></sd-panel></fig><fig id="746"><title></title><label>Figure 3</label><graphic href="https://api.sourcedata.io/file.php?figure_id=746"/><sd-panel panel_id="1089">A <sd-tag id="sdTag313" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>‐<sd-tag id="sdTag314" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag>‐<sd-tag id="sdTag315" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TBC1D5" ext_ids="Q92609" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TBC1D5</sd-tag> expression was induced in <sd-tag id="sdTag106" source="sdapi" category="entity" entity_type="cell_line" role="component" text="T‐REx HeLa" ext_ids="CVCL_D587" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="T-REx-HeLa" ext_tax_ids="" ext_tax_names="" ext_urls="https://identifiers.org/cellosaurus:">T‐REx HeLa</sd-tag> cells for 20 h, lysates were subjected to <sd-tag id="sdTag319" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GST" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GST</sd-tag><sd-tag id="sdTag318" source="sdapi" category="assay" entity_type="" role="" text="pull‐down" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">pull‐down</sd-tag>, −<sd-tag id="sdTag107" source="sdapi" category="entity" entity_type="protein" role="assayed" text="AP2M1" ext_ids="Q96CW1" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="AP2M1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">AP2M1</sd-tag>. <sd-tag id="sdTag114" source="sdapi" category="assay" entity_type="" role="" text="Co‐precipitated" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Co‐precipitated</sd-tag><sd-tag id="sdTag108" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TBC1D5" ext_ids="Q92609" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TBC1D5</sd-tag> was detected with anti−<sd-tag id="sdTag316" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag> antibody.<graphic href="https://api.sourcedata.io/file.php?panel_id=1089"/></sd-panel><sd-panel panel_id="1090">B Control cells and <sd-tag id="sdTag109" source="sdapi" category="entity" entity_type="gene" role="intervention" text="TBC1D5" ext_ids="9779" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">TBC1D5</sd-tag> (shRNA#1) <sd-tag id="sdTag110" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="" ext_tax_names="" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag> cell lysates subjected to <sd-tag id="sdTag111" source="sdapi" category="assay" entity_type="" role="" text="SDS–PAGE" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SDS-PAGE</sd-tag>, subsequently blotted with endogenous antibodies for <sd-tag id="sdTag322" source="sdapi" category="entity" entity_type="subcellular" role="component" text="AP2" ext_ids="GO:0030122" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="AP-2 adaptor complex" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">AP2</sd-tag> subunits <sd-tag id="sdTag112" source="sdapi" category="entity" entity_type="protein" role="assayed" text="AP2A1" ext_ids="O95782" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="AP2A1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">AP2A1</sd-tag> and <sd-tag id="sdTag113" source="sdapi" category="entity" entity_type="protein" role="assayed" text="AP2M1" ext_ids="Q96CW1" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="AP2M1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">AP2M1</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=1090"/></sd-panel><sd-panel panel_id="1091">C <sd-tag id="sdTag119" source="sdapi" category="assay" entity_type="" role="" text="Immunofluorescent staining" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Immunofluorescent staining</sd-tag> of <sd-tag id="sdTag126" source="sdapi" category="entity" entity_type="protein" role="assayed" text="AP2A1" ext_ids="O95782" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="AP2A1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">AP2A1</sd-tag> in <sd-tag id="sdTag125" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="" ext_tax_names="" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag> shRNA control, or shRNA#1 <sd-tag id="sdTag124" source="sdapi" category="entity" entity_type="gene" role="intervention" text="TBC1D5" ext_ids="9779" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">TBC1D5</sd-tag> cells treated with <sd-tag id="sdTag123" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="KU0063974" ext_ids="CHEBI:85572" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Ku-0063794" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">KU0063974</sd-tag> or <sd-tag id="sdTag324" source="sdapi" category="entity" entity_type="molecule" role="component" text="DMSO" ext_ids="CHEBI:28262" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="dimethyl sulfoxide" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DMSO</sd-tag> (6 h).<graphic href="https://api.sourcedata.io/file.php?panel_id=1091"/></sd-panel><sd-panel panel_id="1092">D <sd-tag id="sdTag128" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="" ext_tax_names="" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag> cells stably expressing <sd-tag id="sdTag326" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>‐<sd-tag id="sdTag327" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag>‐<sd-tag id="sdTag129" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ATG9A" ext_ids="Q7Z3C6" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG9A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ATG9A</sd-tag> were fixed and stained for endogenous <sd-tag id="sdTag131" source="sdapi" category="entity" entity_type="protein" role="assayed" text="AP2A1" ext_ids="O95782" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="AP2A1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">AP2A1</sd-tag> and <sd-tag id="sdTag132" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TBC1D5" ext_ids="Q92609" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TBC1D5</sd-tag>. Regions of co‐localization are indicated with arrows.<graphic href="https://api.sourcedata.io/file.php?panel_id=1092"/></sd-panel><sd-panel panel_id="1093">E <sd-tag id="sdTag135" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="" ext_tax_names="" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag> cells stably expressing <sd-tag id="sdTag329" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>‐<sd-tag id="sdTag330" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag>‐<sd-tag id="sdTag242" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ATG9A" ext_ids="Q7Z3C6" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG9A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ATG9A</sd-tag> were starved in <sd-tag id="sdTag140" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> (4 h) or treated with <sd-tag id="sdTag139" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="KU0063794" ext_ids="CHEBI:85572" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Ku-0063794" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">KU0063794</sd-tag> (6 h), subsequently fixed and stained with anti‐<sd-tag id="sdTag141" source="sdapi" category="entity" entity_type="protein" role="assayed" text="AP2A1" ext_ids="O95782" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="AP2A1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">AP2A1</sd-tag> antibody.<graphic href="https://api.sourcedata.io/file.php?panel_id=1093"/></sd-panel><sd-panel panel_id="1094">F <sd-tag id="sdTag142" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="" ext_tax_names="" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag> cells stably expressing <sd-tag id="sdTag143" source="sdapi" category="entity" entity_type="protein" role="reporter" text="mCherry" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">mCherry</sd-tag>‐<sd-tag id="sdTag144" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ATG9A" ext_ids="Q7Z3C6" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG9A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ATG9A</sd-tag> were treated with <sd-tag id="sdTag145" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="KU0063794" ext_ids="CHEBI:85572" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Ku-0063794" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">KU0063794</sd-tag> (6 h), lysed in <sd-tag id="sdTag146" source="sdapi" category="assay" entity_type="" role="" text="co‐immunoprecipitation" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">co‐immunoprecipitation</sd-tag> buffer. Lysates from <sd-tag id="sdTag335" source="sdapi" category="entity" entity_type="molecule" role="component" text="DMSO" ext_ids="CHEBI:28262" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="dimethyl sulfoxide" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DMSO</sd-tag>‐ or <sd-tag id="sdTag336" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="KU0063794" ext_ids="CHEBI:85572" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Ku-0063794" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">KU0063794</sd-tag>‐treated cells were split and equal volumes were incubated with <sd-tag id="sdTag148" 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>‐Trap beads or GFP‐Trap beads, or agarose as a negative control, overnight at 4°C. Precipitated proteins were analyzed by <sd-tag id="sdTag337" source="sdapi" category="assay" entity_type="" role="" text="SDS–PAGE" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SDS-PAGE</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=1094"/></sd-panel></fig><fig id="747"><title></title><label>Figure 4</label><graphic href="https://api.sourcedata.io/file.php?figure_id=747"/><sd-panel panel_id="1095">A <sd-tag id="sdTag152" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="" ext_tax_names="" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag> cells stably expressing <sd-tag id="sdTag340" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>‐<sd-tag id="sdTag341" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag>‐<sd-tag id="sdTag154" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ATG9A" ext_ids="Q7Z3C6" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG9A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ATG9A</sd-tag> transfected with siRNA targeting <sd-tag id="sdTag155" source="sdapi" category="entity" entity_type="gene" role="intervention" text="AP2A1" ext_ids="160" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="AP2A1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">AP2A1</sd-tag> (40 nM), or siRNA AllStar control (40 nM) were treated with <sd-tag id="sdTag156" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="KU0063794" ext_ids="CHEBI:85572" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Ku-0063794" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">KU0063794</sd-tag> or <sd-tag id="sdTag342" source="sdapi" category="entity" entity_type="molecule" role="component" text="DMSO" ext_ids="CHEBI:28262" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="dimethyl sulfoxide" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DMSO</sd-tag> (6 h) 4 days post‐transfection, fixed, and stained with anti‐<sd-tag id="sdTag343" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag> antibody.<graphic href="https://api.sourcedata.io/file.php?panel_id=1095"/></sd-panel><sd-panel panel_id="1096">B Co‐localization of <sd-tag id="sdTag346" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>‐<sd-tag id="sdTag347" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag>‐<sd-tag id="sdTag160" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ATG9A" ext_ids="Q7Z3C6" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG9A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ATG9A</sd-tag> with <sd-tag id="sdTag161" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TGN46" ext_ids="O43493" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TGOLN2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TGN46</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=1096"/></sd-panel><sd-panel panel_id="1097">C Lysates of <sd-tag id="sdTag165" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="" ext_tax_names="" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag> cells transfected with siRNA targeting <sd-tag id="sdTag166" source="sdapi" category="entity" entity_type="gene" role="intervention" text="AP2A1" ext_ids="160" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="AP2A1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">AP2A1</sd-tag>, or siRNA control AllStars analyzed by <sd-tag id="sdTag167" source="sdapi" category="assay" entity_type="" role="" text="SDS–PAGE" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SDS-PAGE</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=1097"/></sd-panel><sd-panel panel_id="1098">D <sd-tag id="sdTag173" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="" ext_tax_names="" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag> cells stably expressing <sd-tag id="sdTag174" source="sdapi" category="entity" entity_type="protein" role="reporter" text="mCherry" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">mCherry</sd-tag>−<sd-tag id="sdTag175" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ATG9" ext_ids="Q7Z3C6" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG9A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ATG9</sd-tag> were transiently transfected with <sd-tag id="sdTag176" 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="sdTag177" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Dynamin 2" ext_ids="P50570" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="DNM2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">Dynamin 2</sd-tag> or GFP−<sd-tag id="sdTag352" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Dynamin 2" ext_ids="1785" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="DNM2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Dynamin 2</sd-tag> (K44A), 20 h post−transfection cells were fixed and with anti−<sd-tag id="sdTag180" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TBC1D5" ext_ids="Q92609" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TBC1D5</sd-tag> antibody. <sd-tag id="sdTag354" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ATG9A" ext_ids="Q7Z3C6" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG9A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ATG9A</sd-tag> and <sd-tag id="sdTag355" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Dynamin 2" ext_ids="P50570" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="DNM2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">Dynamin 2</sd-tag>co−localization is indicated by arrows.<graphic href="https://api.sourcedata.io/file.php?panel_id=1098"/></sd-panel></fig><fig id="748"><title></title><label>Figure 5</label><graphic href="https://api.sourcedata.io/file.php?figure_id=748"/><sd-panel panel_id="1099">A <sd-tag id="sdTag184" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="" ext_tax_names="" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag> cells stably expressing <sd-tag id="sdTag356" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>−<sd-tag id="sdTag357" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag>−<sd-tag id="sdTag186" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ATG9A" ext_ids="Q7Z3C6" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG9A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ATG9A</sd-tag> treated with <sd-tag id="sdTag358" source="sdapi" category="entity" entity_type="molecule" role="component" text="DMSO" ext_ids="CHEBI:28262" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="dimethyl sulfoxide" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DMSO</sd-tag> or <sd-tag id="sdTag187" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="Dynasore" ext_ids="5717066" norm_text="" ext_dbs="PubChem" in_caption="True" ext_names="Dynasore" ext_tax_ids="" ext_tax_names="" ext_urls="https://pubchem.ncbi.nlm.nih.gov/compound/">Dynasore</sd-tag> (100 μM) 15 min, fixed and <sd-tag id="sdTag360" source="sdapi" category="assay" entity_type="" role="" text="immunostained" ext_ids="BAO_0002426" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunofluorescent labeling" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunostained</sd-tag> with anti−<sd-tag id="sdTag359" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag> and anti−<sd-tag id="sdTag189" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TBC1D5" ext_ids="Q92609" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TBC1D5</sd-tag> antibodies.<graphic href="https://api.sourcedata.io/file.php?panel_id=1099"/></sd-panel><sd-panel panel_id="1100">B Quantification of <sd-tag id="sdTag192" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ATG9A" ext_ids="Q7Z3C6" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG9A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ATG9A</sd-tag>and <sd-tag id="sdTag193" source="sdapi" category="entity" entity_type="protein" role="assayed" text="AP2A1" ext_ids="O95782" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="AP2A1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">AP2A1</sd-tag>co‐localization from experiment in (A), pictures from 3 independent experiments were analyzed and co‐localization quantified, statistics is calculated as described in Supplementary Materials and Methods.<graphic href="https://api.sourcedata.io/file.php?panel_id=1100"/></sd-panel><sd-panel panel_id="1101">C RGB‐line profile across the vesicle indicated with arrow in (B); red-<sd-tag id="sdTag200" source="sdapi" category="entity" entity_type="protein" role="assayed" text="AP2A1" ext_ids="O95782" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="AP2A1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">AP2A1</sd-tag>; green-<sd-tag id="sdTag198" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ATG9A" ext_ids="Q7Z3C6" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG9A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ATG9A</sd-tag>; blue-<sd-tag id="sdTag199" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TBC1D5" ext_ids="Q92609" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TBC1D5</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=1101"/></sd-panel><sd-panel panel_id="1102">D Quantification of <sd-tag id="sdTag202" source="sdapi" category="entity" entity_type="protein" role="assayed" text="ATG9A" ext_ids="Q7Z3C6" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG9A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ATG9A</sd-tag> and <sd-tag id="sdTag223" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="AP2" ext_ids="GO:0030122" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="AP-2 adaptor complex" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">AP2</sd-tag> co‐localization. Cells were starved in <sd-tag id="sdTag205" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> media (45 min) or pretreated with <sd-tag id="sdTag206" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="Dynasore" ext_ids="5717066" norm_text="" ext_dbs="PubChem" in_caption="True" ext_names="Dynasore" ext_tax_ids="" ext_tax_names="" ext_urls="https://pubchem.ncbi.nlm.nih.gov/compound/">Dynasore</sd-tag> (15 min) and subsequently starved for 45 min in combination with <sd-tag id="sdTag370" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="Dynasore" ext_ids="5717066" norm_text="" ext_dbs="PubChem" in_caption="True" ext_names="Dynasore" ext_tax_ids="" ext_tax_names="" ext_urls="https://pubchem.ncbi.nlm.nih.gov/compound/">Dynasore</sd-tag>. Control cells were treated with <sd-tag id="sdTag371" source="sdapi" category="entity" entity_type="molecule" role="component" text="DMSO" ext_ids="CHEBI:28262" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="dimethyl sulfoxide" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DMSO</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=1102"/></sd-panel><sd-panel panel_id="1103">E Quantification of <sd-tag id="sdTag207" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9GZQ8///Q9BXW4///Q9H492" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3B///MAP1LC3C///MAP1LC3A" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag> puncta in cells stably expressing <sd-tag id="sdTag373" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>‐<sd-tag id="sdTag374" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag>‐<sd-tag id="sdTag209" source="sdapi" category="entity" entity_type="protein" role="component" text="ATG9A" ext_ids="Q7Z3C6" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG9A" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">ATG9A</sd-tag>, treated and presented as in (D). 100 cells were quantified in 3 independent experiments.<graphic href="https://api.sourcedata.io/file.php?panel_id=1103"/></sd-panel><sd-panel panel_id="1104">F <sd-tag id="sdTag213" source="sdapi" category="entity" entity_type="cell_line" role="component" text="T‐REx HeLa" ext_ids="CVCL_D587" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="T-REx-HeLa" ext_tax_ids="" ext_tax_names="" ext_urls="https://identifiers.org/cellosaurus:">T‐REx HeLa</sd-tag> cells were transfected with siRNA Control oligo or siRNA oligo#1 targeting (40 nM). 72 h post−transfection, expression of <sd-tag id="sdTag383" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>‐<sd-tag id="sdTag384" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Flag" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Flag</sd-tag>−<sd-tag id="sdTag216" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TBC1D5" ext_ids="Q92609" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TBC1D5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TBC1D5</sd-tag> was induced with <sd-tag id="sdTag218" source="sdapi" category="entity" entity_type="molecule" role="component" text="doxycycline" ext_ids="CHEBI:50845" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="doxycycline" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">doxycycline</sd-tag> (50 ng/ml), and 96 h post−transfection cells were treated with <sd-tag id="sdTag217" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="Dynasore" ext_ids="5717066" norm_text="" ext_dbs="PubChem" in_caption="True" ext_names="Dynasore" ext_tax_ids="" ext_tax_names="" ext_urls="https://pubchem.ncbi.nlm.nih.gov/compound/">Dynasore</sd-tag> for 15 min, or pretreated with <sd-tag id="sdTag385" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="Dynasore" ext_ids="5717066" norm_text="" ext_dbs="PubChem" in_caption="True" ext_names="Dynasore" ext_tax_ids="" ext_tax_names="" ext_urls="https://pubchem.ncbi.nlm.nih.gov/compound/">Dynasore</sd-tag> and subsequently starved in combination with <sd-tag id="sdTag386" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="Dynasore" ext_ids="5717066" norm_text="" ext_dbs="PubChem" in_caption="True" ext_names="Dynasore" ext_tax_ids="" ext_tax_names="" ext_urls="https://pubchem.ncbi.nlm.nih.gov/compound/">Dynasore</sd-tag> for additional 45 min. Cells were lysed in <sd-tag id="sdTag222" source="sdapi" category="assay" entity_type="" role="" text="co‐immunoprecipitation" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">co‐immunoprecipitation</sd-tag> buffer, and lysates were incubated with <sd-tag id="sdTag381" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Myc</sd-tag> antibody or M2 antibody coupled with agarose overnight at 4°C. Beads were washed 3 times with incubation buffer and subjected to <sd-tag id="sdTag382" source="sdapi" category="assay" entity_type="" role="" text="SDS‐PAGE" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SDS‐PAGE</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=1104"/></sd-panel></fig></article>