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<article doi="10.1038/emboj.2009.369" abstract="In addition to mitochondria, BCL-2 is located at the endoplasmic reticulum (ER) where it is a constituent of several distinct complexes. Here, we identify the BCL-2-interacting protein at the ER, nutrient-deprivation autophagy factor-1 (NAF-1)-a bitopic integral membrane protein whose defective expression underlies the aetiology of the neurodegenerative disorder Wolfram syndrome 2 (WFS2). NAF-1 contains a two iron-two sulphur coordinating domain within its cytosolic region, which is necessary, but not sufficient for interaction with BCL-2. NAF-1 is displaced from BCL-2 by the ER-restricted BH3-only protein BIK and contributes to regulation of BIK-initiated autophagy, but not BIK-dependent activation of caspases. Similar to BCL-2, NAF-1 is found in association with the inositol 1,4,5-triphosphate receptor and is required for BCL-2-mediated depression of ER Ca(2+) stores. During nutrient deprivation as a physiological stimulus of autophagy, BCL-2 is known to function through inhibition of the autophagy effector and tumour suppressor Beclin 1. NAF-1 is required in this pathway for BCL-2 at the ER to functionally antagonize Beclin 1-dependent autophagy. Thus, NAF-1 is a BCL-2-associated co-factor that targets BCL-2 for antagonism of the autophagy pathway at the ER."><fig id="2570"><title></title><label>Figure 1</label><graphic href="https://api.sourcedata.io/file.php?figure_id=2570"/><sd-panel panel_id="7329"><div>(A) BH3‐only <sd-tag id="sdTag28" source="sdapi" category="entity" entity_type="gene" role="intervention" text="BIK" ext_ids="638" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="BIK" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">BIK</sd-tag> displaces a subset of <sd-tag id="sdTag29" source="sdapi" category="entity" entity_type="protein" role="assayed" text="BCL‐2" ext_ids="P10415" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">BCL‐2</sd-tag> complexes at the <sd-tag id="sdTag30" source="sdapi" category="entity" entity_type="subcellular" role="component" text="ER" ext_ids="GO:0005783" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="endoplasmic reticulum" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">ER</sd-tag>. Purified <sd-tag id="sdTag1" source="sdapi" category="entity" entity_type="subcellular" role="component" text="LM" ext_ids="GO:0005789" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="endoplasmic reticulum membrane" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">LM</sd-tag> from <sd-tag id="sdTag3" source="sdapi" category="entity" entity_type="cell_line" role="component" text="H1299" ext_ids="CVCL_0060" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="NCI-H1299" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">H1299</sd-tag> <sd-tag id="sdTag4" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>‐<sd-tag id="sdTag42" source="sdapi" category="entity" entity_type="gene" role="component" text="BCL‐2" ext_ids="596" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">BCL‐2</sd-tag>b5 cells either mock infected or infected with <sd-tag id="sdTag6" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Ad" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Ad</sd-tag>‐<sd-tag id="sdTag7" source="sdapi" category="entity" entity_type="gene" role="intervention" text="BIK" ext_ids="638" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="BIK" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">BIK</sd-tag>b5 was subjected to cross‐linking with <sd-tag id="sdTag8" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="BMH" ext_ids="18179261" norm_text="" ext_dbs="PubChem" in_caption="True" ext_names="bismaleimidohexane" ext_tax_ids="" ext_tax_names="" ext_urls="https://pubchem.ncbi.nlm.nih.gov/compound/">BMH</sd-tag> and visualized by <sd-tag id="sdTag285" source="sdapi" category="assay" entity_type="" role="" text="immunoblot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblot</sd-tag>. Asterisks (*) denote <sd-tag id="sdTag9" source="sdapi" category="entity" entity_type="protein" role="assayed" text="BCL‐2" ext_ids="P10415" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">BCL‐2</sd-tag> cross‐linked products, which are displaced by <sd-tag id="sdTag10" source="sdapi" category="entity" entity_type="gene" role="intervention" text="BIK" ext_ids="638" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="BIK" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">BIK</sd-tag>. Bold arrow denotes region containing cross‐linked <sd-tag id="sdTag11" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NAF‐1" ext_ids="Q8N5K1" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">NAF‐1</sd-tag>/<sd-tag id="sdTag12" source="sdapi" category="entity" entity_type="protein" role="assayed" text="BCL‐2" ext_ids="P10415" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">BCL‐2</sd-tag>.</div><graphic href="https://api.sourcedata.io/file.php?panel_id=7329"/></sd-panel><sd-panel panel_id="7332">(D) Endogenous <sd-tag id="sdTag20" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NAF‐1" ext_ids="Q8N5K1" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">NAF‐1</sd-tag> cross‐links with <sd-tag id="sdTag16" source="sdapi" category="entity" entity_type="protein" role="assayed" text="BCL‐2" ext_ids="P10415" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">BCL‐2</sd-tag> at the <sd-tag id="sdTag17" source="sdapi" category="entity" entity_type="subcellular" role="component" text="ER" ext_ids="GO:0005783" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="endoplasmic reticulum" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">ER</sd-tag>. <sd-tag id="sdTag21" source="sdapi" category="entity" entity_type="subcellular" role="component" text="LM" ext_ids="GO:0005789" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="endoplasmic reticulum membrane" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">LM</sd-tag> purified from <sd-tag id="sdTag22" source="sdapi" category="entity" entity_type="cell_line" role="component" text="H1299" ext_ids="CVCL_0060" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="NCI-H1299" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">H1299</sd-tag> <sd-tag id="sdTag292" source="sdapi" category="entity" entity_type="protein" role="reporter" text="neo" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">neo</sd-tag> and <sd-tag id="sdTag23" 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="sdTag43" source="sdapi" category="entity" entity_type="gene" role="intervention" text="BCL‐2" ext_ids="596" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">BCL‐2</sd-tag>b5 cells were subjected to cross‐linking by <sd-tag id="sdTag25" source="sdapi" category="entity" entity_type="molecule" role="component" text="BMH" ext_ids="18179261" norm_text="" ext_dbs="PubChem" in_caption="True" ext_names="bismaleimidohexane" ext_tax_ids="" ext_tax_names="" ext_urls="https://pubchem.ncbi.nlm.nih.gov/compound/">BMH</sd-tag> and immunoprecipitation with anti‐<sd-tag id="sdTag26" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NAF‐1" ext_ids="Q8N5K1" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">NAF‐1</sd-tag> antibody. Precipitates were analysed by <sd-tag id="sdTag291" source="sdapi" category="assay" entity_type="" role="" text="immunoblot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblot</sd-tag> with anti‐<sd-tag id="sdTag27" source="sdapi" category="entity" entity_type="protein" role="assayed" text="BCL‐2" ext_ids="P10415" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">BCL‐2</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=7332"/></sd-panel><sd-panel panel_id="7333">(E) <sd-tag id="sdTag34" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NAF‐1" ext_ids="Q8N5K1" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">NAF‐1</sd-tag> is displaced from <sd-tag id="sdTag33" source="sdapi" category="entity" entity_type="protein" role="assayed" text="BCL‐2" ext_ids="P10415" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">BCL‐2</sd-tag> by BH3‐only <sd-tag id="sdTag35" source="sdapi" category="entity" entity_type="gene" role="intervention" text="BIK" ext_ids="638" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="BIK" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">BIK</sd-tag>. <sd-tag id="sdTag36" source="sdapi" category="entity" entity_type="cell_line" role="component" text="H1299" ext_ids="CVCL_0060" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="NCI-H1299" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">H1299</sd-tag> <sd-tag id="sdTag37" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>‐<sd-tag id="sdTag44" source="sdapi" category="entity" entity_type="gene" role="component" text="BCL‐2" ext_ids="596" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">BCL‐2</sd-tag>b5 cells were either mock infected or infected with <sd-tag id="sdTag39" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Ad" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Ad</sd-tag>‐<sd-tag id="sdTag40" source="sdapi" category="entity" entity_type="gene" role="intervention" text="BIK" ext_ids="638" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="BIK" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">BIK</sd-tag>. Purified <sd-tag id="sdTag41" source="sdapi" category="entity" entity_type="subcellular" role="component" text="LM" ext_ids="GO:0005789" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="endoplasmic reticulum membrane" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">LM</sd-tag> was treated as in (D).<graphic href="https://api.sourcedata.io/file.php?panel_id=7333"/></sd-panel></fig><fig id="2571"><title></title><label>Figure 2</label><graphic href="https://api.sourcedata.io/file.php?figure_id=2571"/><sd-panel panel_id="7334">(A) <sd-tag id="sdTag49" 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="sdTag52" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NAF‐1" ext_ids="Q8N5K1" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">NAF‐1</sd-tag> and <sd-tag id="sdTag53" source="sdapi" category="entity" entity_type="protein" role="assayed" text="BCL‐2" ext_ids="P10415" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">BCL‐2</sd-tag>. Lysates from <sd-tag id="sdTag50" source="sdapi" category="entity" entity_type="cell_line" role="component" text="SK‐Mel5 cells" ext_ids="CVCL_0527" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="SK-MEL-5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">SK‐Mel5 cells</sd-tag> were collected and immunoprecipitation was performed with anti‐<sd-tag id="sdTag54" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NAF‐1" ext_ids="Q8N5K1" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">NAF‐1</sd-tag> antibody. The precipitate was subjected to <sd-tag id="sdTag51" source="sdapi" category="assay" entity_type="" role="" text="immunoblotting" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotting</sd-tag> with anti‐<sd-tag id="sdTag55" source="sdapi" category="entity" entity_type="protein" role="assayed" text="BCL‐2" ext_ids="P10415" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">BCL‐2</sd-tag> and anti‐<sd-tag id="sdTag56" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NAF‐1" ext_ids="Q8N5K1" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">NAF‐1</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=7334"/></sd-panel><sd-panel panel_id="7335">(B) <sd-tag id="sdTag58" source="sdapi" category="entity" entity_type="cell_line" role="component" text="H1299" ext_ids="CVCL_0060" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="NCI-H1299" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">H1299</sd-tag> cells were fixed and double stained with anti‐<sd-tag id="sdTag59" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NAF‐1" ext_ids="Q8N5K1" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">NAF‐1</sd-tag> and anti‐<sd-tag id="sdTag60" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Calnexin" ext_ids="P27824" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CANX" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">Calnexin</sd-tag> or anti‐<sd-tag id="sdTag61" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Cytochrome c" ext_ids="P08574" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CYC1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">Cytochrome c</sd-tag> antibodies. Scale bar represents 10 μm.<graphic href="https://api.sourcedata.io/file.php?panel_id=7335"/></sd-panel><sd-panel panel_id="7336">(C) <sd-tag id="sdTag65" 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="sdTag66" source="sdapi" category="entity" entity_type="gene" role="component" text="NAF‐1" ext_ids="493856" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">NAF‐1</sd-tag>‐<sd-tag id="sdTag73" 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> and <sd-tag id="sdTag67" 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="sdTag63" source="sdapi" category="entity" entity_type="protein" role="assayed" text="BCL‐2" ext_ids="P10415" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">BCL‐2</sd-tag>b5. Lysates from <sd-tag id="sdTag68" source="sdapi" category="entity" entity_type="cell_line" role="component" text="H1299" ext_ids="CVCL_0060" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="NCI-H1299" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">H1299</sd-tag> <sd-tag id="sdTag69" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>‐<sd-tag id="sdTag64" source="sdapi" category="entity" entity_type="gene" role="component" text="BCL‐2" ext_ids="596" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">BCL‐2</sd-tag>b5 cells infected with <sd-tag id="sdTag70" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Ad" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Ad</sd-tag>‐<sd-tag id="sdTag71" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NAF‐1" ext_ids="Q8N5K1" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">NAF‐1</sd-tag>‐<sd-tag id="sdTag72" 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> were collected and treated as in (A).<graphic href="https://api.sourcedata.io/file.php?panel_id=7336"/></sd-panel><sd-panel panel_id="7337">(D) Mutations in the CDGSH iron‐binding domain of <sd-tag id="sdTag83" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NAF‐1" ext_ids="Q8N5K1" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">NAF‐1</sd-tag> interfere with <sd-tag id="sdTag84" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NAF‐1" ext_ids="Q8N5K1" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">NAF‐1</sd-tag> binding to <sd-tag id="sdTag85" source="sdapi" category="entity" entity_type="protein" role="assayed" text="BCL‐2" ext_ids="P10415" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">BCL‐2</sd-tag>. <sd-tag id="sdTag86" source="sdapi" category="entity" entity_type="cell_line" role="component" text="H1299" ext_ids="CVCL_0060" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="NCI-H1299" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">H1299</sd-tag> <sd-tag id="sdTag87" 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="sdTag75" source="sdapi" category="entity" entity_type="gene" role="component" text="BCL‐2" ext_ids="596" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">BCL‐2</sd-tag>b5 cells were infected with either <sd-tag id="sdTag88" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Ad" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Ad</sd-tag>‐<sd-tag id="sdTag76" source="sdapi" category="entity" entity_type="gene" role="component" text="rtTA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">rtTA</sd-tag>, <sd-tag id="sdTag89" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Ad" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Ad</sd-tag>‐<sd-tag id="sdTag77" source="sdapi" category="entity" entity_type="gene" role="intervention" text="NAF‐1" ext_ids="493856" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">NAF‐1</sd-tag>‐<sd-tag id="sdTag90" 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>, or <sd-tag id="sdTag91" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Ad" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Ad</sd-tag>‐<sd-tag id="sdTag78" source="sdapi" category="entity" entity_type="gene" role="intervention" text="NAF‐1" ext_ids="493856" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">NAF‐1</sd-tag>‐mut‐<sd-tag id="sdTag92" 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> (C99S C101S C110S H114Q). Lysates were treated as in (A). Densitometric analysis was performed using Scion Image software to quantify expression and co‐precipitated levels of <sd-tag id="sdTag93" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NAF‐1" ext_ids="Q8N5K1" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">NAF‐1</sd-tag>‐<sd-tag id="sdTag94" 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> and <sd-tag id="sdTag95" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NAF‐1" ext_ids="Q8N5K1" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">NAF‐1</sd-tag>‐mut‐<sd-tag id="sdTag96" 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>. Graph depicts the ratio of co‐precipitated protein to expression level.<graphic href="https://api.sourcedata.io/file.php?panel_id=7337"/></sd-panel><sd-panel panel_id="7338">(E) A functional CDGSH iron‐binding domain is necessary, but not sufficient for the interaction between the cytosolic domains of <sd-pretag id="sdPretag14388680794770" parent-tag-id="309">NAF‐1</sd-pretag> and <sd-pretag id="sdPretag14388680335270" parent-tag-id="308">BCL‐2</sd-pretag>. HA‐<sd-pretag id="sdPretag14388680335271" parent-tag-id="308">BCL‐2</sd-pretag> ΔTM was in vitro translated in <sd-tag id="sdTag295" source="sdapi" category="entity" entity_type="organism" role="component" text="rabbit" ext_ids="9986" norm_text="" ext_dbs="NCBI taxon" in_caption="True" ext_names="rabbit" ext_tax_ids="9986" ext_tax_names="Oryctolagus cuniculus" ext_urls="http://www.ncbi.nlm.nih.gov/taxonomy/">rabbit</sd-tag> reticulocyte lysate and equivalent aliquots were added to each GST pull‐down reaction. GST‐fusion proteins used were GST alone, GST‐NAF1‐C, GST‐NAF1‐C‐mut (C99S C101S C110S H114Q), and GST‐MitoNEET‐C. The proteins were detected using anti‐HA and anti‐GST<graphic href="https://api.sourcedata.io/file.php?panel_id=7338"/></sd-panel></fig><fig id="2572"><title></title><label>Figure 3</label><graphic href="https://api.sourcedata.io/file.php?figure_id=2572"/><sd-panel panel_id="7339">(A) <sd-tag id="sdTag122" source="sdapi" category="entity" entity_type="cell_line" role="component" text="H1299" ext_ids="CVCL_0060" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="NCI-H1299" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">H1299</sd-tag> <sd-tag id="sdTag296" source="sdapi" category="entity" entity_type="protein" role="reporter" text="neo" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">neo</sd-tag> and <sd-tag id="sdTag123" 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="sdTag121" source="sdapi" category="entity" entity_type="gene" role="intervention" text="BCL‐2" ext_ids="596" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">BCL‐2</sd-tag>b5 cells treated with control (CTRL) or <sd-tag id="sdTag124" source="sdapi" category="entity" entity_type="gene" role="intervention" text="NAF‐1" ext_ids="493856" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">NAF‐1</sd-tag> shRNA were either mock infected or infected with <sd-tag id="sdTag125" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Ad" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Ad</sd-tag>‐<sd-tag id="sdTag126" source="sdapi" category="entity" entity_type="gene" role="intervention" text="BIK" ext_ids="638" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="BIK" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">BIK</sd-tag> in the absence or presence of 50 μM <sd-tag id="sdTag127" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="zVAD‐fmk" ext_ids="5737" norm_text="" ext_dbs="PubChem" in_caption="True" ext_names="Z-VAD(OMe)-FMK" ext_tax_ids="" ext_tax_names="" ext_urls="https://pubchem.ncbi.nlm.nih.gov/compound/">zVAD‐fmk</sd-tag>. Cell lysates were analysed by <sd-tag id="sdTag297" source="sdapi" category="assay" entity_type="" role="" text="immunoblot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblot</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=7339"/></sd-panel><sd-panel panel_id="7340">(B) Caspase activity was measured using the fluorescent substrate <sd-tag id="sdTag134" source="sdapi" category="assay" entity_type="" role="" text="DEVD‐AMC" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">DEVD‐AMC</sd-tag>. The results represent the average±s.d. of three independent experiments.<graphic href="https://api.sourcedata.io/file.php?panel_id=7340"/></sd-panel><sd-panel panel_id="7341">(C) Prolonged <sd-tag id="sdTag152" source="sdapi" category="entity" entity_type="gene" role="intervention" text="BIK" ext_ids="638" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="BIK" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">BIK</sd-tag> expression and caspase inhibition induces autophagy, which is enhanced by knockdown of <sd-tag id="sdTag153" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NAF‐1" ext_ids="Q8N5K1" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">NAF‐1</sd-tag>. <sd-tag id="sdTag154" source="sdapi" category="entity" entity_type="cell_line" role="component" text="H1299" ext_ids="CVCL_0060" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="NCI-H1299" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">H1299</sd-tag> <sd-tag id="sdTag299" source="sdapi" category="entity" entity_type="protein" role="reporter" text="neo" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">neo</sd-tag> and <sd-tag id="sdTag155" 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="sdTag149" source="sdapi" category="entity" entity_type="gene" role="intervention" text="BCL‐2" ext_ids="596" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">BCL‐2</sd-tag>b5 cells treated with CTRL or <sd-tag id="sdTag150" source="sdapi" category="entity" entity_type="gene" role="intervention" text="NAF‐1" ext_ids="493856" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">NAF‐1</sd-tag> shRNA were infected with <sd-tag id="sdTag156" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Ad" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Ad</sd-tag>‐<sd-tag id="sdTag151" source="sdapi" category="entity" entity_type="gene" role="intervention" text="BIK" ext_ids="638" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="BIK" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">BIK</sd-tag> in the presence of <sd-tag id="sdTag157" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="zVAD‐fmk" ext_ids="5737" norm_text="" ext_dbs="PubChem" in_caption="True" ext_names="Z-VAD(OMe)-FMK" ext_tax_ids="" ext_tax_names="" ext_urls="https://pubchem.ncbi.nlm.nih.gov/compound/">zVAD‐fmk</sd-tag> for the indicated periods of time. Cell lysates were analysed by <sd-tag id="sdTag298" source="sdapi" category="assay" entity_type="" role="" text="immunoblot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblot</sd-tag>. Levels of <sd-tag id="sdTag300" 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> II were normalized to <sd-tag id="sdTag159" source="sdapi" category="entity" entity_type="protein" role="normalizing" text="actin" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">actin</sd-tag> levels by densitometry analysis. Graph depicts normalized <sd-tag id="sdTag301" 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> II levels of each lane.<graphic href="https://api.sourcedata.io/file.php?panel_id=7341"/></sd-panel></fig><fig id="2573"><title></title><label>Figure 4</label><graphic href="https://api.sourcedata.io/file.php?figure_id=2573"/><sd-panel panel_id="7342">(A) Effect of <sd-tag id="sdTag172" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NAF‐1" ext_ids="Q8N5K1" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">NAF‐1</sd-tag> knockdown on starvation‐induced autophagy. <sd-tag id="sdTag173" source="sdapi" category="entity" entity_type="cell_line" role="component" text="H1299" ext_ids="CVCL_0060" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="NCI-H1299" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">H1299</sd-tag> cells infected with CTRL or <sd-tag id="sdTag171" source="sdapi" category="entity" entity_type="gene" role="intervention" text="NAF‐1" ext_ids="493856" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">NAF‐1</sd-tag> shRNA were starved for 4 h in <sd-tag id="sdTag303" source="sdapi" category="entity" entity_type="undefined" role="component" text="EBSS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">EBSS</sd-tag> with <sd-tag id="sdTag165" 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> (vehicle) or <sd-tag id="sdTag168" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="Baf A1" ext_ids="CHEBI:22689" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="bafilomycin A1" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">Baf A1</sd-tag> (100 nM). Cell lysates were analysed by <sd-tag id="sdTag169" source="sdapi" category="assay" entity_type="" role="" text="immunoblot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblot</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=7342"/></sd-panel><sd-panel panel_id="7343">(B) Representative images of <sd-tag id="sdTag180" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>‐<sd-tag id="sdTag179" source="sdapi" category="entity" entity_type="gene" role="component" text="LC3" ext_ids="440738///81631///84557" norm_text="" ext_dbs="NCBI gene///NCBI gene///NCBI gene" in_caption="True" ext_names="MAP1LC3C///MAP1LC3B///MAP1LC3A" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene/">LC3</sd-tag> staining in <sd-tag id="sdTag183" source="sdapi" category="entity" entity_type="cell_line" role="component" text="H1299" ext_ids="CVCL_0060" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="NCI-H1299" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">H1299</sd-tag> <sd-tag id="sdTag184" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>‐<sd-tag id="sdTag185" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9BXW4///Q9H492///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3C///MAP1LC3A///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag> cells transfected with <sd-tag id="sdTag186" source="sdapi" category="entity" entity_type="gene" role="component" text="LUC" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">LUC</sd-tag> or <sd-tag id="sdTag181" source="sdapi" category="entity" entity_type="gene" role="intervention" text="NAF‐1" ext_ids="493856" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">NAF‐1</sd-tag> siRNA, untreated and starved. Scale bar represents 10 μm.<graphic href="https://api.sourcedata.io/file.php?panel_id=7343"/></sd-panel><sd-panel panel_id="7344">(C) Quantification of autophagy is expressed as the percentage of <sd-tag id="sdTag188" 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="sdTag189" source="sdapi" category="entity" entity_type="gene" role="component" text="LC3" ext_ids="440738///81631///84557" norm_text="" ext_dbs="NCBI gene///NCBI gene///NCBI gene" in_caption="True" ext_names="MAP1LC3C///MAP1LC3B///MAP1LC3A" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene/">LC3</sd-tag>‐expressing cells displaying punctate <sd-tag id="sdTag190" 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="sdTag191" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9H492///Q9BXW4///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3A///MAP1LC3C///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag>. A minimum of 100 cells per sample were counted; results represent the average±s.d. of three independent experiments. Cell lysates were analysed by <sd-tag id="sdTag192" source="sdapi" category="assay" entity_type="" role="" text="immunoblot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblot</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=7344"/></sd-panel></fig><fig id="2574"><title></title><label>Figure 5</label><graphic href="https://api.sourcedata.io/file.php?figure_id=2574"/><sd-panel panel_id="7345">(A) <sd-tag id="sdTag204" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NAF‐1" ext_ids="Q8N5K1" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">NAF‐1</sd-tag> contributes to the interaction between <sd-tag id="sdTag205" source="sdapi" category="entity" entity_type="protein" role="assayed" text="BCL‐2" ext_ids="P10415" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">BCL‐2</sd-tag> and <sd-tag id="sdTag206" source="sdapi" category="entity" entity_type="gene" role="component" text="Beclin 1" ext_ids="8678" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="BECN1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Beclin 1</sd-tag>. <sd-tag id="sdTag207" source="sdapi" category="entity" entity_type="cell_line" role="component" text="H1299" ext_ids="CVCL_0060" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="NCI-H1299" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">H1299</sd-tag> <sd-tag id="sdTag208" 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="sdTag200" source="sdapi" category="entity" entity_type="gene" role="component" text="BCL‐2" ext_ids="596" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">BCL‐2</sd-tag>b5 cells were transfected with <sd-tag id="sdTag209" 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="sdTag201" source="sdapi" category="entity" entity_type="gene" role="component" text="Beclin 1" ext_ids="8678" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="BECN1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Beclin 1</sd-tag> and either <sd-tag id="sdTag210" source="sdapi" category="entity" entity_type="gene" role="component" text="LUC" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">LUC</sd-tag> or <sd-tag id="sdTag202" source="sdapi" category="entity" entity_type="gene" role="intervention" text="NAF‐1" ext_ids="493856" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">NAF‐1</sd-tag> siRNA. Cells were lysed and subjected to immunoprecipitation with anti‐<sd-tag id="sdTag203" source="sdapi" category="entity" entity_type="protein" role="assayed" text="BCL‐2" ext_ids="P10415" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">BCL‐2</sd-tag> antibody. Precipitates were subjected to analysis by <sd-tag id="sdTag211" source="sdapi" category="assay" entity_type="" role="" text="immunoblot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblot</sd-tag> using anti‐<sd-tag id="sdTag212" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Beclin 1" ext_ids="Q14457" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="BECN1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">Beclin 1</sd-tag> and anti‐<sd-tag id="sdTag213" source="sdapi" category="entity" entity_type="gene" role="component" text="BCL‐2" ext_ids="596" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">BCL‐2</sd-tag>. All lanes are derived from the same gel and of the same exposure. Thin white lines indicate where lanes have been removed.<graphic href="https://api.sourcedata.io/file.php?panel_id=7345"/></sd-panel><sd-panel panel_id="7346">(B) Loss of <sd-tag id="sdTag310" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NAF‐1" ext_ids="Q8N5K1" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">NAF‐1</sd-tag> prevents <sd-tag id="sdTag311" source="sdapi" category="entity" entity_type="protein" role="assayed" text="BCL‐2" ext_ids="P10415" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">BCL‐2</sd-tag>b5 from antagonizing starvation‐induced autophagy. <sd-tag id="sdTag222" source="sdapi" category="entity" entity_type="cell_line" role="assayed" text="H1299" ext_ids="CVCL_0060" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="NCI-H1299" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">H1299</sd-tag> GFP‐<sd-tag id="sdTag312" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LC3" ext_ids="Q9BXW4///Q9H492///Q9GZQ8" norm_text="" ext_dbs="Uniprot///Uniprot///Uniprot" in_caption="True" ext_names="MAP1LC3C///MAP1LC3A///MAP1LC3B" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">LC3</sd-tag> and <sd-tag id="sdTag308" source="sdapi" category="entity" entity_type="gene" role="intervention" text="BCL‐2" ext_ids="596" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">BCL‐2</sd-tag>b5/<sd-tag id="sdTag221" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>‐<sd-tag id="sdTag226" source="sdapi" category="entity" entity_type="gene" role="component" text="LC3" ext_ids="440738///81631///84557" norm_text="" ext_dbs="NCBI gene///NCBI gene///NCBI gene" in_caption="True" ext_names="MAP1LC3C///MAP1LC3B///MAP1LC3A" ext_tax_ids="9606///9606///9606" ext_tax_names="Homo sapiens///Homo sapiens///Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene////http://www.ncbi.nlm.nih.gov/gene/">LC3</sd-tag> cells were transfected with either <sd-tag id="sdTag218" source="sdapi" category="entity" entity_type="gene" role="component" text="LUC" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">LUC</sd-tag> or <sd-tag id="sdTag309" source="sdapi" category="entity" entity_type="gene" role="intervention" text="NAF‐1" ext_ids="493856" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">NAF‐1</sd-tag> siRNA and starved for 4 h. Cells were analysed as in Figure 4C.<graphic href="https://api.sourcedata.io/file.php?panel_id=7346"/></sd-panel><sd-panel panel_id="7347">(C) Representative electron micrographs of <sd-tag id="sdTag237" source="sdapi" category="entity" entity_type="cell_line" role="assayed" text="H1299" ext_ids="CVCL_0060" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="NCI-H1299" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">H1299</sd-tag> <sd-tag id="sdTag313" source="sdapi" category="entity" entity_type="protein" role="reporter" text="neo" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">neo</sd-tag> and <sd-tag id="sdTag238" 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="sdTag232" source="sdapi" category="entity" entity_type="gene" role="intervention" text="BCL‐2" ext_ids="596" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">BCL‐2</sd-tag>b5 cells treated with either <sd-tag id="sdTag239" source="sdapi" category="entity" entity_type="gene" role="component" text="LUC" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">LUC</sd-tag> or <sd-tag id="sdTag233" source="sdapi" category="entity" entity_type="gene" role="intervention" text="NAF‐1" ext_ids="493856" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">NAF‐1</sd-tag> siRNA, with or without subsequent starvation. Scale bar represents 10 μm. Cell lysates were analysed by <sd-tag id="sdTag235" source="sdapi" category="assay" entity_type="" role="" text="immunoblot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblot</sd-tag>. All lanes are derived from the same gel and of the same exposure. Thin white line indicates where lanes have been removed. Autophagy levels observed by <sd-tag id="sdTag315" source="sdapi" category="assay" entity_type="" role="" text="electron microscopy" ext_ids="BAO_0000449" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="electron microscopy" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">electron microscopy</sd-tag> (C) were quantified and expressed as either the percentage of cells containing <sd-tag id="sdTag236" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="autophagic vacuoles" ext_ids="GO:0005776" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="autophagic vacuole" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">autophagic vacuoles</sd-tag> (D) or the number of <sd-tag id="sdTag231" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="autophagosomes" ext_ids="GO:0005776" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="autophagic vacuole" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">autophagosomes</sd-tag> per cell (E). A minimum of 100 cells per sample were counted; results represent the average±s.e.m.<graphic href="https://api.sourcedata.io/file.php?panel_id=7347"/></sd-panel></fig><fig id="2575"><title></title><label>Figure 6</label><graphic href="https://api.sourcedata.io/file.php?figure_id=2575"/><sd-panel panel_id="7348">(A) <sd-tag id="sdTag245" source="sdapi" category="entity" entity_type="cell_line" role="component" text="H1299" ext_ids="CVCL_0060" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="NCI-H1299" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">H1299</sd-tag> <sd-tag id="sdTag317" source="sdapi" category="entity" entity_type="protein" role="reporter" text="neo" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">neo</sd-tag> and <sd-tag id="sdTag246" 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="sdTag247" source="sdapi" category="entity" entity_type="gene" role="intervention" text="BCL‐2" ext_ids="596" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">BCL‐2</sd-tag>b5 cells treated with either CTRL or <sd-tag id="sdTag248" source="sdapi" category="entity" entity_type="gene" role="intervention" text="NAF‐1" ext_ids="493856" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">NAF‐1</sd-tag> shRNA were loaded with <sd-tag id="sdTag249" source="sdapi" category="assay" entity_type="" role="" text="Fura‐2AM" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Fura‐2AM</sd-tag>, and <sd-tag id="sdTag250" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="ER" ext_ids="GO:0005783" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="endoplasmic reticulum" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">ER</sd-tag> Ca2+ stores were measured as the difference in cytoplasmic Ca2+ concentration before and after addition of <sd-tag id="sdTag252" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="TG" ext_ids="CHEBI:9516" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="thapsigargin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">TG</sd-tag> (2 μM). Shown are representative traces of <sd-tag id="sdTag256" source="sdapi" category="assay" entity_type="" role="" text="Fura‐2AM" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Fura‐2AM</sd-tag> fluorescence measured at 340/380 nm excitation wavelength ratio at 510 nm wavelength emission. Arrow indicates time at which <sd-tag id="sdTag257" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="TG" ext_ids="CHEBI:9516" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="thapsigargin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">TG</sd-tag> was added, delta values indicate <sd-tag id="sdTag258" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="TG" ext_ids="CHEBI:9516" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="thapsigargin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">TG</sd-tag>‐sensitive <sd-tag id="sdTag259" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="ER" ext_ids="GO:0005783" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="endoplasmic reticulum" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">ER</sd-tag> <sd-tag id="sdTag255" source="sdapi" category="entity" entity_type="molecule" role="assayed" text="Ca2+" ext_ids="CHEBI:22984" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="calcium atom" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">Ca2+</sd-tag> stores.<graphic href="https://api.sourcedata.io/file.php?panel_id=7348"/></sd-panel><sd-panel panel_id="7349">(B) Differences in <sd-tag id="sdTag318" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="ER" ext_ids="GO:0005783" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="endoplasmic reticulum" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">ER</sd-tag> <sd-tag id="sdTag262" source="sdapi" category="entity" entity_type="molecule" role="assayed" text="Ca2+" ext_ids="CHEBI:22984" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="calcium atom" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">Ca2+</sd-tag> stores are shown as the average±s.e.m. of three independent experiments as described in (A).<graphic href="https://api.sourcedata.io/file.php?panel_id=7349"/></sd-panel><sd-panel panel_id="7350">(C) <sd-tag id="sdTag274" 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="sdTag275" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NAF‐1" ext_ids="Q8N5K1" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CISD2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">NAF‐1</sd-tag> and endogenous <sd-tag id="sdTag288" source="sdapi" category="entity" entity_type="protein" role="assayed" text="IP3 receptor type I" ext_ids="Q14643" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ITPR1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">IP3 receptor type I</sd-tag>. Lysate from <sd-tag id="sdTag276" source="sdapi" category="entity" entity_type="cell_line" role="component" text="H1299" ext_ids="CVCL_0060" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="NCI-H1299" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">H1299</sd-tag> <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="sdTag279" source="sdapi" category="entity" entity_type="gene" role="component" text="BCL‐2" ext_ids="596" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="BCL2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">BCL‐2</sd-tag>b5 cells was collected and immunoprecipitation was performed with anti‐<sd-tag id="sdTag283" source="sdapi" category="entity" entity_type="protein" role="assayed" text="IP3R1" ext_ids="Q14643" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ITPR1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">IP3R1</sd-tag> antibody. The resulting precipitate was analysed by <sd-tag id="sdTag278" source="sdapi" category="assay" entity_type="" role="" text="immunoblot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblot</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=7350"/></sd-panel></fig></article>