MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy. Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy. EMBO J. (0261-4189) 2011 imex curation Strappazzon F., Vietri-Rudan M., Campello S., Nazio F., Florenzano F., Fimia GM., Piacentini M., Levine B., Cecconi F. Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. crosslink cross-linking study crosslink western blot western blot Immuno blot Immuno blot Strappazzon F., Vietri-Rudan M., Campello S., Nazio F., Florenzano F., Fimia GM., Piacentini M., Levine B., Cecconi F. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy. Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy. EMBO J. (0261-4189) 2011 imex curation Strappazzon F., Vietri-Rudan M., Campello S., Nazio F., Florenzano F., Fimia GM., Piacentini M., Levine B., Cecconi F. Only protein-protein interactions in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip predetermined predetermined participant predetermined Strappazzon F., Vietri-Rudan M., Campello S., Nazio F., Florenzano F., Fimia GM., Piacentini M., Levine B., Cecconi F. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy. Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy. EMBO J. (0261-4189) 2011 imex curation Strappazzon F., Vietri-Rudan M., Campello S., Nazio F., Florenzano F., Fimia GM., Piacentini M., Levine B., Cecconi F. Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Strappazzon F., Vietri-Rudan M., Campello S., Nazio F., Florenzano F., Fimia GM., Piacentini M., Levine B., Cecconi F. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy. Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy. EMBO J. (0261-4189) 2011 imex curation Strappazzon F., Vietri-Rudan M., Campello S., Nazio F., Florenzano F., Fimia GM., Piacentini M., Levine B., Cecconi F. Only protein-protein interactions in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Strappazzon F., Vietri-Rudan M., Campello S., Nazio F., Florenzano F., Fimia GM., Piacentini M., Levine B., Cecconi F. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy. Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy. EMBO J. (0261-4189) 2011 imex curation Strappazzon F., Vietri-Rudan M., Campello S., Nazio F., Florenzano F., Fimia GM., Piacentini M., Levine B., Cecconi F. Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip predetermined predetermined participant predetermined Strappazzon F., Vietri-Rudan M., Campello S., Nazio F., Florenzano F., Fimia GM., Piacentini M., Levine B., Cecconi F. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy. Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy. EMBO J. (0261-4189) 2011 imex curation Strappazzon F., Vietri-Rudan M., Campello S., Nazio F., Florenzano F., Fimia GM., Piacentini M., Levine B., Cecconi F. Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Strappazzon F., Vietri-Rudan M., Campello S., Nazio F., Florenzano F., Fimia GM., Piacentini M., Levine B., Cecconi F. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy. Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy. EMBO J. (0261-4189) 2011 imex curation Strappazzon F., Vietri-Rudan M., Campello S., Nazio F., Florenzano F., Fimia GM., Piacentini M., Levine B., Cecconi F. Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti bait coip anti bait coimmunoprecipitation anti bait coip predetermined predetermined participant predetermined Strappazzon F., Vietri-Rudan M., Campello S., Nazio F., Florenzano F., Fimia GM., Piacentini M., Levine B., Cecconi F. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy. Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy. EMBO J. (0261-4189) 2011 imex curation Strappazzon F., Vietri-Rudan M., Campello S., Nazio F., Florenzano F., Fimia GM., Piacentini M., Levine B., Cecconi F. Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Strappazzon F., Vietri-Rudan M., Campello S., Nazio F., Florenzano F., Fimia GM., Piacentini M., Levine B., Cecconi F. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy. Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy. EMBO J. (0261-4189) 2011 imex curation Strappazzon F., Vietri-Rudan M., Campello S., Nazio F., Florenzano F., Fimia GM., Piacentini M., Levine B., Cecconi F. Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Strappazzon F., Vietri-Rudan M., Campello S., Nazio F., Florenzano F., Fimia GM., Piacentini M., Levine B., Cecconi F. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation becn1_human Beclin-1 Coiled-coil myosin-like BCL2-interacting protein Protein GT197 BECN1 GT197 protein protein human Homo sapiens Human MEGSKTSNNSTMQVSFVCQRCSQPLKLDTSFKILDRVTIQELTAPLLTTAQAKPGETQEEETNSGEEPFIETPRQDGVSRRFIPPARMMSTESANSFTLIGEASDGGTMENLSRRLKVTGDLFDIMSGQTDVDHPLCEECTDTLLDQLDTQLNVTENECQNYKRCLEILEQMNEDDSEQLQMELKELALEEERLIQELEDVEKNRKIVAENLEKVQAEAERLDQEEAQYQREYSEFKRQQLELDDELKSVENQMRYAQTQLDKLKKTNVFNATFHIWHSGQFGTINNFRLGRLPSVPVEWNEINAAWGQTVLLLHALANKMGLKFQRYRLVPYGNHSYLESLTDKSKELPLYCSGGLRFFWDNKFDHAMVAFLDCVQQFKEEVEKGETRFCLPYRMDVEKGKIEDTGGSGGSYSIKTQFNSEEQWTKALKFMLTNLKWGLAWVSSQFYNK DIP protein Q14457 original sequence version: 95 DIP protein Q14457 original sequence version: 156 DIP protein Q14457 original sequence version: 122 DIP protein Q14457 original sequence version: 120 DIP protein Q14457 original sequence version: 133 DIP protein Q14457 original sequence version: 141 DIP protein Q14457 original sequence version: 106 ABF0C2DD7087473C amra1_human Activating molecule in BECN1-regulated autophagy protein 1 AMBRA1 DCAF3 DDB1- and CUL4-associated factor 3 KIAA1736 protein protein human Homo sapiens Human MKVVPEKNAVRILWGRERGARAMGAQRLLQELVEDKTRWMKWEGKRVELPDSPRSTFLLAFSPDRTLLASTHVNHNIYITEVKTGKCVHSLIGHRRTPWCVTFHPTISGLIASGCLDGEVRIWDLHGGSESWFTDSNNAIASLAFHPTAQLLLIATANEIHFWDWSRREPFAVVKTASEMERVRLVRFDPLGHYLLTAIVNPSNQQGDDEPEIPIDGTELSHYRQRALLQSQPVRRTPLLHNFLHMLSSRSSGIQVGEQSTVQDSATPSPPPPPPQPSTERPRTSAYIRLRQRVSYPTAECCQHLGILCLCSRCSGTRVPSLLPHQDSVPPASARATTPSFSFVQTEPFHPPEQASSTQQDQGLLNRPSAFSTVQSSTAGNTLRNLSLGPTRRSLGGPLSSHPSRYHREIAPGLTGSEWTRTVLSLNSRSEAESMPPPRTSASSVSLLSVLRQQEGGSQASVYTSATEGRGFPASGLATESDGGNGSSQNNSGSIRHELQCDLRRFFLEYDRLQELDQSLSGEAPQTQQAQEMLNNNIESERPGPSHQPTPHSSENNSNLSRGHLNRCRACHNLLTFNNDTLRWERTTPNYSSGEASSSWQVPSSFESVPSSGSQLPPLERTEGQTPSSSRLELSSSASPQEERTVGVAFNQETGHWERIYTQSSRSGTVSQEALHQDMPEESSEEDSLRRRLLESSLISLSRYDGAGSREHPIYPDPARLSPAAYYAQRMIQYLSRRDSIRQRSMRYQQNRLRSSTSSSSSDNQGPSVEGTDLEFEDFEDNGDRSRHRAPRNARMSAPSLGRFVPRRFLLPEYLPYAGIFHERGQPGLATHSSVNRVLAGAVIGDGQSAVASNIANTTYRLQWWDFTKFDLPEISNASVNVLVQNCKIYNDASCDISADGQLLAAFIPSSQRGFPDEGILAVYSLAPHNLGEMLYTKRFGPNAISVSLSPMGRYVMVGLASRRILLHPSTEHMVAQVFRLQQAHGGETSMRRVFNVLYPMPADQRRHVSINSARWLPEPGLGLAYGTNKGDLVICRPEALNSGVEYYWDQLNETVFTVHSNSRSSERPGTSRATWRTDRDMGLMNAIGLQPRNPATSVTSQGTQTLALQLQNAETQTEREVPEPGTAASGPGEGEGSEYGASGEDALSRIQRLMAEGGMTAVVQREQSTTMASMGGFGNNIIVSHRIHRSSQTGTEPGAAHTSSPQPSTSRGLLPEAGQLAERGLSPRTASWDQPGTPGREPTQPTLPSSSPVPIPVSLPSAEGPTLHCELTNNNHLLDGGSSRGDAAGPRGEPRNR DIP protein Q9C0C7 original sequence version: 104 DIP protein Q9C0C7 original sequence version: 96 44FE9CDFFFE6811E bcl2_human Apoptosis regulator Bcl-2 BCL2 protein protein human Homo sapiens Human MAHAGRTGYDNREIVMKYIHYKLSQRGYEWDAGDVGAAPPGAAPAPGIFSSQPGHTPHPAASRDPVARTSPLQTPAAPGAAAGPALSPVPPVVHLTLRQAGDDFSRRYRRDFAEMSSQLHLTPFTARGRFATVVEELFRDGVNWGRIVAFFEFGGVMCVESVNREMSPLVDNIALWMTEYLNRHLHTWIQDNGGWDAFVELYGPSMRPLFDFSWLSLKTLLSLALVGACITLGAYLGHK DIP protein P10415 original sequence version: 148 DIP protein P10415 original sequence version: 125 DIP protein P10415 original sequence version: 151 DIP protein P10415 original sequence version: 204 DIP protein P10415 original sequence version: 149 3C49F2B714DC9CCB ambra1-bcl2 6 BCL2 12 immunostaining immunostaining predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell AMBRA1 11 immunostaining immunostaining predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization f1c homomint mint ambra1-bcl2-1 8 AMBRA1 11 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level BCL2 12 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f1d homomint mint ambra1-bcl2-2 5 AMBRA1 11 anti tag western anti tag western blot anti tag western predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell myc tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain BCL2 12 anti tag western anti tag western blot anti tag western predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association f1b homomint mint bcl2-ambra1 5 BCL2 12 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined AMBRA1 11 anti tag western anti tag western blot anti tag western predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell myc tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain physical association physical association homomint sf1 mint becn1-ambra1 5 Beclin 10 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined AMBRA1 11 anti tag western anti tag western blot anti tag western predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell myc tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association homomint mint f7b f7d becn1-bcl2-1 5 Beclin 10 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined BCL2 12 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell P10415:p.Ser70Glu mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain S E Tue Jul 12 10:54:22 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 17:22:54 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser70glu' Tue Jun 30 19:51:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser70Glu' P10415:p.Thr69Glu mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain T E Tue Jul 12 10:23:43 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 18:33:07 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr69glu' Tue Jun 30 15:56:26 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr69Glu' P10415:p.Ser87Glu mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain S E Tue Jul 12 10:23:42 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 16:17:29 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser87glu' Tue Jun 30 17:11:55 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser87Glu' physical association physical association f2b homomint mint ambra1-bcl2-3 7 BCL2 12 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell AMBRA1 11 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell myc tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association f2c homomint mint ambra1-bcl2-4 7 BCL2 12 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level Beclin 10 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level AMBRA1 11 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level association association sf2e homomint mint becn1-ambra1-1 7 Beclin 10 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell undefined mutation mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 15:05:39 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation disrupting' 88-150 AMBRA1 11 anti tag western anti tag western blot anti tag western predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell myc tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association f7c homomint mint ambra1-bcl2-5 3 BCL2 12 tag fluorescence tag visualisation by fluorescence tag fluorescence predetermined predetermined participant predetermined electron donor electron donor neutral component neutral component over-expressed over expressed level over-expressed living cell living cell AMBRA1 11 tag fluorescence tag visualisation by fluorescence tag fluorescence predetermined predetermined participant predetermined electron acceptor electron acceptor neutral component neutral component over-expressed over expressed level over-expressed living cell living cell fusion protein fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined proximity proximity close-contact colocalization neighbourhood interaction f1e homomint mint ambra1-bcl2-6 1 AMBRA1 11 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level BCL2 12 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level covalent binding covalent binding f3d homomint mint To consolidate these data, we decided to carry out a mitochondrial crosslink before performing a subcellular fractionation experiment in HEK293 cells grown in normal or starvation conditions. As illustrated in Figure 3D, when cells are not fixed, BCL-2 can be detected as a 26-kDa band (appropriate molecular weight) in normal condition, and this band decreases following autophagy induction. However, after cell fixation, this band is not detectable, whereas a band of B170 kDa is detectable. The 170-kDa band may corresponds most likely to a complex including at least endogenous AMBRA1 and BCL-2, thus confirming the interaction. ambra1-bcl2-7 9 AMBRA1 11 immunostaining immunostaining predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell BCL2 12 immunostaining immunostaining predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization homomint mint sf4b sf4c becn1-ambra1-2 2 Beclin 10 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep over-expressed over expressed level over-expressed flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined AMBRA1 11 anti tag western anti tag western blot anti tag western predetermined predetermined participant predetermined unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep over-expressed over expressed level over-expressed myc tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association homomint mint sf6b ambra1-bcl2-8 4 AMBRA1 11 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level BCL2 12 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f3c homomint mint We performed an immunoprecipitation of AMBRA1 in the mitochondrial fractions, and we were able to appreciate the binding with endogenous BCL-2 in normal conditions. In contrast, this binding was disrupted after autophagy induction (Figure 3C). To consolidate these data, we decided to carry out a mitochondrial crosslink before performing a subcellular fractionation experiment in HEK293 cells grown in normal or starvation conditions. As illustrated in Figure 3D, when cells are not fixed, BCL-2 can be detected as a 26-kDa band (appropriate molecular weight) in normal condition, and this band decreases following autophagy induction. However, after cell fixation, this band is not detectable, whereas a band of B170 kDa is detectable. The 170-kDa band may corresponds most likely to a complex including at least endogenous AMBRA1 and BCL-2, thus confirming the interaction.