IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ Involvement of Nrf2 in proteasome inhibition-mediated induction of ORP150 in thyroid cancer cells. Involvement of Nrf2 in proteasome inhibition-mediated induction of ORP150 in thyroid cancer cells. Oncotarget 2016 imex curation Zong ZH., Du ZX., Zhang HY., Li C., An MX., Li S., Yao HB., Wang HQ. wanghq_doctor@hotmail.com 27-Mar-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-8305c human-8305c Human thyroid anaplastic carcinoma, undifferentiated Human 8305c 8305-C cell pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Zong ZH., Du ZX., Zhang HY., Li C., An MX., Li S., Yao HB., Wang HQ. MG132 treatment wanghq_doctor@hotmail.com Accepted 2018-FEB-19 AT 10:58 GMT AT 10:58 GMT by MGT Oncotarget 2016 Only protein-protein interactions imex curation imex curation I created another exp and re-distribuited interactions accordingly. Involvement of Nrf2 in proteasome inhibition-mediated induction of ORP150 in thyroid cancer cells. Involvement of Nrf2 in proteasome inhibition-mediated induction of ORP150 in thyroid cancer cells. Oncotarget 2016 imex curation Zong ZH., Du ZX., Zhang HY., Li C., An MX., Li S., Yao HB., Wang HQ. wanghq_doctor@hotmail.com 27-Mar-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-8305c human-8305c Human thyroid anaplastic carcinoma, undifferentiated Human 8305c 8305-C cell ch-ip chromatin immunoprecipitation assay ch-ip primer specific pcr primer specific pcr Zong ZH., Du ZX., Zhang HY., Li C., An MX., Li S., Yao HB., Wang HQ. MG132 treatment wanghq_doctor@hotmail.com Accepted 2018-FEB-19 AT 10:58 GMT AT 10:58 GMT by MGT Oncotarget 2016 Only protein-protein interactions imex curation imex curation I created another exp and re-distribuited interactions accordingly. Involvement of Nrf2 in proteasome inhibition-mediated induction of ORP150 in thyroid cancer cells. Involvement of Nrf2 in proteasome inhibition-mediated induction of ORP150 in thyroid cancer cells. Oncotarget 2016 imex curation Zong ZH., Du ZX., Zhang HY., Li C., An MX., Li S., Yao HB., Wang HQ. wanghq_doctor@hotmail.com 27-Mar-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-8305c human-8305c Human thyroid anaplastic carcinoma, undifferentiated Human 8305c 8305-C cell ch-ip chromatin immunoprecipitation assay ch-ip q-pcr quantitative pcr q-pcr Zong ZH., Du ZX., Zhang HY., Li C., An MX., Li S., Yao HB., Wang HQ. MG132 treatment wanghq_doctor@hotmail.com Accepted 2018-FEB-19 AT 10:58 GMT AT 10:58 GMT by MGT Oncotarget 2016 Only protein-protein interactions imex curation imex curation I created another exp and re-distribuited interactions accordingly. atf4_human Cyclic AMP-dependent transcription factor ATF-4 Activating transcription factor 4 CREB2 ATF4 TXREB Cyclic AMP-responsive element-binding protein 2 Tax-responsive enhancer element-binding protein 67 protein protein human Homo sapiens Human MTEMSFLSSEVLVGDLMSPFDQSGLGAEESLGLLDDYLEVAKHFKPHGFSSDKAKAGSSEWLAVDGLVSPSNNSKEDAFSGTDWMLEKMDLKEFDLDALLGIDDLETMPDDLLTTLDDTCDLFAPLVQETNKQPPQTVNPIGHLPESLTKPDQVAPFTFLQPLPLSPGVLSSTPDHSFSLELGSEVDITEGDRKPDYTAYVAMIPQCIKEEDTPSDNDSGICMSPESYLGSPQHSPSTRGSPNRSLPSPGVLCGSARPKPYDPPGEKMVAAKVKGEKLDKKLKKMEQNKTAATRYRQKKRAEQEALTGECKELEKKNEALKERADSLAKEIQYLKDLIEEVRKARGKKRVP 3BBB7379DC3B0D07 DIP protein P18848 original sequence version: 181 DIP protein P18848 original sequence version: 117 No Uniprot Update and Caution removed " The protein could not be merged with EBI-12907955 because of some conflicts with the protein sequence (features which cannot be shifted)." -EBI-12907955 no longer exists. 3BBB7379DC3B0D07 orp150_human_gene HYOU1 gene gene human Homo sapiens Human nf2l2_human Nuclear factor erythroid 2-related factor 2 NFE2L2 NRF2 Nuclear factor, erythroid derived 2, like 2 HEBP1 protein protein human Homo sapiens Human MMDLELPPPGLPSQQDMDLIDILWRQDIDLGVSREVFDFSQRRKEYELEKQKKLEKERQEQLQKEQEKAFFAQLQLDEETGEFLPIQPAQHIQSETSGSANYSQVAHIPKSDALYFDDCMQLLAQTFPFVDDNEVSSATFQSLVPDIPGHIESPVFIATNQAQSPETSVAQVAPVDLDGMQQDIEQVWEELLSIPELQCLNIENDKLVETTMVPSPEAKLTEVDNYHFYSSIPSMEKEVGNCSPHFLNAFEDSFSSILSTEDPNQLTVNSLNSDATVNTDFGDEFYSAFIAEPSISNSMPSPATLSHSLSELLNGPIDVSDLSLCKAFNQNHPESTAEFNDSDSGISLNTSPSVASPEHSVESSSYGDTLLGLSDSEVEELDSAPGSVKQNGPKTPVHSSGDMVQPLSPSQGQSTHVHDAQCENTPEKELPVSPGHRKTPFTKDKHSSRLEAHLTRDELRAKALHIPFPVEKIINLPVVDFNEMMSKEQFNEAQLALIRDIRRRGKNKVAAQNCRKRKLENIVELEQDLDHLKDEKEKLLKEKGENDKSLHLLKKQLSTLYLEVFSMLRDEDGKPYSPSEYSLQQTRDGNVFLVPKSKKPDVKKN DIP protein Q16236 original sequence version: 80 DIP protein Q16236 original sequence version: 103 DIP protein Q16236 original sequence version: 148 DIP protein Q16236 original sequence version: 102 DIP protein Q16236-1 original sequence version: 113 99FAFD811B6C1416 atf4-nrf2-1 1 4 unspecified role unspecified role neutral component neutral component living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 6 unspecified role unspecified role neutral component neutral component living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level proximity proximity close-contact colocalization neighbourhood interaction Fig. 4D Duolink PLA confirmed endogenous binding between ATF4 and Nrf2 in 8305C cells, which was enhanced by MG132 exposure (Figure 4D). MG132 nrf2-porp150-1 2 4 unspecified role unspecified role ancillary ancillary knock-down knock down knock-down 5 unspecified role unspecified role prey prey living cell living cell promoter-region-243/+53 promoter-region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association Fig.4C Nrf2 directly transactivated the ORP150 gene by direct binding with the -421/-307 region. Nrf2 also indirectly activated OPR150 transcription via facilitating recruitment of ATF4 to the -243/+53 region. nfe2l2-hyou1-1 2 5 unspecified role unspecified role prey prey living cell living cell promoter-region-421/-307 promoter-region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association Fig.4C Nrf2 directly transactivated the ORP150 gene by direct binding with the -421/-307 region. Nrf2 also indirectly activated OPR150 transcription via facilitating recruitment of ATF4 to the -243/+53 region. nfe2l2-hyou1-4 3 4 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 5 unspecified role unspecified role prey prey living cell living cell promoter-region-ERSE promoter-region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role ancillary ancillary knock-down knock down knock-down association association Fig.4F Knockdown of Nrf2 expression by siNrf2 significantly decreased the binding of ATF4 to the ERSE element on the ORP150 promoter in 8305C cells (Figure ​4F), indicating that Nrf2 might stimulate the recruitment of ATF4 to the ORP150 promoter. nfe2l2-atf4-1 2 4 unspecified role unspecified role prey prey partially purified partially purified 5 unspecified role unspecified role prey prey partially purified partially purified promoter-region-243/+53 promoter-region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role bait bait partially purified partially purified association association Fig.4E re-ChIP by using ATF4 and Nrf2 antibodies sequentially demonstrated that after the ATF4 first ChIP enrichment of Nrf2 binding was observed in the ORP150 promoter at -243/+53 region in 8305C cells (Figure 4E), indicating that Nrf2 and ATF4 were concurrently bound to the ORP150 promoter at this region. On the contrary, Nrf2 binding was not observed in the ORP150 promoter at -421/-307 region after the ATF4 first ChIP (Figure 4E). nfe2l2-hyou1-5 3 5 unspecified role unspecified role prey prey living cell living cell promoter-region-421/-307 promoter-region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined promoter-region-243/+53 sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association Fig.3A Nrf2 directly transactivated the ORP150 gene by direct binding with the -421/-307 region. Nrf2 also indirectly activated OPR150 transcription via facilitating recruitment of ATF4 to the -243/+53 region. atf4-hyou1-2 3 5 unspecified role unspecified role prey prey living cell living cell promoter-region-243/+53 promoter-region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association Fig.4A ChIP analysis using an antibody against ATF4 demonstrated that MG132 increased recruitment of ATF4 to the -243/+53 region of the ORP150 gene promoter in 8305C cells (Figure 4A).