Fig. 3

TRAF3IP2 promotes the progression of NONO-TFE3 tRCC by activating NOTCH signaling pathway. A-B HEK 293 T cells were co-transfected with TRAF3IP2 plasmid and HES1 reporter plasmid, and the luciferase activity was determined using a dual luciferase reporter assay after 48 h. C UOK109 cells were lysed, then CoIP and reverse CoIP were performed with anti-TRAF3IP2 antibody, anti-NOTCH1 antibody or normal rabbit IgG. D 786-O cells were transfected with Flag-TRAF3IP2 or HA-NOTCH1, then CoIP assay was performed with anti-Flag antibody, anti-HA antibody or normal rabbit IgG. E UOK109 cells were transfected with Flag-TRAF3IP2, HA-NICD1 and V5-NECD1, then CoIP assay was performed with anti-Flag antibody. F Nuclear and cytosolic extracts of UOK109 cells were isolated after treatment with DAPT, and the nuclear translocation of TRAF3IP2 and NICD1 were analyzed by western blot. G-I UOK109 or 786-O cells were transfected with indicated lentivirus, then CoIP assay was performed with anti-Flag antibody or anti-RBPJ antibody. J ChIP assays showed TRAF3IP2 and RBPJ binding to the promoter region of NOTCH1 target genes. K-L UOK109 cells were treated with DAPT or transfected with shTRAF3IP2, then ChIP assay was performed with anti-TRAF3IP2 antibody or anti-NOTCH1 antibody. M–N The mRNA level of target genes of NOTCH1 was detected in UOK109 cells and 786-O cells transfected indicated lentivirus. The data are presented as the mean ± SD, *P < 0.05, **P < 0.01, ***P < 0.001