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 http://mint.bio.uniroma2.it/mint lu-2008-15 human-bmsc human Human bmsc Bone Marrow Stromal cells ch-ip chromatin immunoprecipitation assay ch-ip primer specific pcr primer specific pcr Lu C., Chen JQ., Zhou GP., Wu SH., Guan YF., Yuan CS., Lu C., Chen JQ., Zhou GP., Wu SH., Guan YF., Yuan CS. luchao8009@163.com Nucleic acids research 2008 imex curation Only protein-protein interactions imex curation lu-2008-16 in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Lu C., Chen JQ., Zhou GP., Wu SH., Guan YF., Yuan CS., Lu C., Chen JQ., Zhou GP., Wu SH., Guan YF., Yuan CS. luchao8009@163.com Nucleic acids research 2008 imex curation Only protein-protein interactions imex curation lu-2008-14 human-bmsc human Human bmsc Bone Marrow Stromal cells anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Lu C., Chen JQ., Zhou GP., Wu SH., Guan YF., Yuan CS., Lu C., Chen JQ., Zhou GP., Wu SH., Guan YF., Yuan CS. luchao8009@163.com Nucleic acids research 2008 imex curation Only protein-protein interactions imex curation lu-2008-17 in vitro In vitro emsa electrophoretic mobility shift assay Gel retardation assay band shift band shift Gel retardation assay emsa weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Lu C., Chen JQ., Zhou GP., Wu SH., Guan YF., Yuan CS., Lu C., Chen JQ., Zhou GP., Wu SH., Guan YF., Yuan CS. luchao8009@163.com Nucleic acids research 2008 imex curation Only protein-protein interactions imex curation lu-2008-18 human-bmsc human Human bmsc Bone Marrow Stromal cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Lu C., Chen JQ., Zhou GP., Wu SH., Guan YF., Yuan CS., Lu C., Chen JQ., Zhou GP., Wu SH., Guan YF., Yuan CS. luchao8009@163.com Nucleic acids research 2008 imex curation Only protein-protein interactions imex curation e2f1_human Transcription factor E2F1 Retinoblastoma-associated protein 1 Retinoblastoma-binding protein 3 PBR3 E2F1 RBBP3 pRB-binding protein E2F-1 protein protein human Homo sapiens Human MALAGAPAGGPCAPALEALLGAGALRLLDSSQIVIISAAQDASAPPAPTGPAAPAAGPCDPDLLLFATPQAPRPTPSAPRPALGRPPVKRRLDLETDHQYLAESSGPARGRGRHPGKGVKSPGEKSRYETSLNLTTKRFLELLSHSADGVVDLNWAAEVLKVQKRRIYDITNVLEGIQLIAKKSKNHIQWLGSHTTVGVGGRLEGLTQDLRQLQESEQQLDHLMNICTTQLRLLSEDTDSQRLAYVTCQDLRSIADPAEQMVMVIKAPPETQLQAVDSSENFQISLKSKQGPIDVFLCPEETVGGISPGKTPSQEVTSEEENRATDSATIVSPPPSSPPSSLTTDPSQSLLSLEQEPLLSRMGSLRAPVDEDRLSPLVAADSLLEHVREDFSGLLPEEFISLSPPHEALDYHFGLEEGEGIRDLFDCDFGDLTPLDF dbloh_human_gene Diablo homolog, mitochondrial DIABLO SMAC gene gene human Homo sapiens Human pawr_human PRKC apoptosis WT1 regulator protein Prostate apoptosis response 4 protein PAWR PAR4 protein protein human Homo sapiens Human MATGGYRTSSGLGGSTTDFLEEWKAKREKMRAKQNPPGPAPPGGGSSDAAGKPPAGALGTPAAAAANELNNNLPGGAPAAPAVPGPGGVNCAVGSAMLTRAAPGPRRSEDEPPAASASAAPPPQRDEEEPDGVPEKGKSSGPSARKGKGQIEKRKLREKRRSTGVVNIPAAECLDEYEDDEAGQKERKREDAITQQNTIQNEAVNLLDPGSSYLLQEPPRTVSGRYKSTTSVSEEDVSSRYSRTDRSGFPRYNRDANVSGTLVSSSTLEKKIEDLEKEVVRERQENLRLVRLMQDKEEMIGKLKEEIDLLNRDLDDIEDENEQLKQENKTLLKVVGQLTR diablo_human_e2f1-site_dna-probe Human Diablo gene promoter transcription factor E2F1 binding BS3 site DNA probe dna deoxyribonucleic acid DNA deoxyribonucleic acid dna DNA deoxyribonucleic acid human Homo sapiens Human GCCCCGCGCGGAAGTT e2f1-1 3450314 N/A 3450319 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3450320 unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level -200 to -193bp sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false f5 Chromatin immunoprecipitated with anti-Par-4 antibody or anti-E2F1 antibody was subjected to PCR using primers representing BS2 (−542 to −535 bp relative to ATG) site, another previously reported E2F1-binding site. No DNA could be amplified from the chromatins, (data not shown), demonstrating that Par-4 and E2F1 specifically bind to the BS3 (−200 to −193 bp relative to ATG) site on the Smac promoter. Glutamate diablo-lpar4-1 3450314 N/A 3450321 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3450320 unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level -200 to -193bp sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false f5 Chromatin immunoprecipitated with anti-Par-4 antibody or anti-E2F1 antibody was subjected to PCR using primers representing BS2 (−542 to −535 bp relative to ATG) site, another previously reported E2F1-binding site. No DNA could be amplified from the chromatins, (data not shown), demonstrating that Par-4 and E2F1 specifically bind to the BS3 (−200 to −193 bp relative to ATG) site on the Smac promoter. Glutamate diablo_human_e2f1_site_dna_probe-e2f1-3 3450315 N/A 3450322 unspecified role unspecified role bait bait purified purified biotin tag biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3450319 unspecified role unspecified role prey prey partially purified partially purified human-bmsc human Human bmsc Bone Marrow Stromal cells physical association physical association false false false f4b Indeed, we found that Par-4 and E2F1 existed in nuclear extracts from glutamate-treated cells (Figure 4A, lane 1; Figure 4B, lane 1). Glutamate diablo_human_e2f1_site_dna_probe-e2f1-1 3450315 N/A 3450322 unspecified role unspecified role bait bait purified purified biotin tag biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3450321 unspecified role unspecified role prey prey partially purified partially purified human-bmsc human Human bmsc Bone Marrow Stromal cells physical association physical association false false false f4a Indeed, we found that Par-4 and E2F1 existed in nuclear extracts from glutamate-treated cells (Figure 4A, lane 1; Figure 4B, lane 1). Glutamate e2f1-10 3450316 N/A 3450321 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell myc tag myc tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region binding region region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false N/A 3450319 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 false false false f6 As shown in Figure 3, Par-4 co-immunoprecipitated with E2F1 protein in nuclear extracts from the glutamate-treated hBMSCs, indicating the direct interaction between Par-4 and E2F1. By contrast, in nuclear extracts from the hBMSCs without the treatment of glutamate, Par-4 failed to associate with E2F1. diablo_human_e2f1_site_dna_probe-e2f1-2 3450317 N/A 3450319 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level human-bmsc human Human bmsc Bone Marrow Stromal cells N/A 3450322 unspecified role unspecified role bait bait purified purified chemical synthesis Chemical synthesis (Chemical synthesis) physical association physical association false false false f4c The neutralizing antibody against E2F1 caused disappearance of both the faster and the slower migrating complex, thus indicating the presence of E2F1 in both the complexes (Figure 4C, lane 5). These results supported that it was E2F1, not Par-4, in Par-4-E2F1 proteins complex that directly bound to Smac promoter, and that Par-4 might indirectly exert its transcriptional control on Smac gene though interacting with E2F1. The BS3 E2F1-binding site was also used as a probe to perform EMSA with nuclear extracts from the hBMSCs without treatment of glutamate. However, no DNA–protein complex was found (Figure 4C, lane 7). Glutamate e2f1-9 3450318 N/A 3450321 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3450319 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false f3 As shown in Figure 3, Par-4 co-immunoprecipitated with E2F1 protein in nuclear extracts from the glutamate-treated hBMSCs, indicating the direct interaction between Par-4 and E2F1. By contrast, in nuclear extracts from the hBMSCs without the treatment of glutamate, Par-4 failed to associate with E2F1. Glutamate