Fig. 6

HNRNPK suppresses TRAF3IP2 via H3K9me3 through recruiting SETDB1. A 786-O cells transfected with Flag-SETDB1 and UOK109 cells were lysed, then CoIP assay was performed with anti-SETDB1 antibody, anti-Flag antibody or normal rabbit IgG. B-C HEK 293 T cells were transfected with SETDB1 truncations or HNRNPK truncations, then CoIP assay was performed. D-E UOK109 cells and 786-O cells were transfected with indicated lentivirus, then ChIP assay was performed with anti-H3 antibody or anti- H3K9me3 antibody. F–H UOK109 cells and 786-O cells were transfected with indicated lentivirus or treatment with indicated inhibitors, then the mRNA level of TRAF3IP2 was analyzed. I-K The mRNA and protein levels of TRAF3IP2 and SETDB1 were detected in UOK109 cells and 786-O cells transfected shSETDB1. L Schematic illustration of dCas9-based system to deliver the exogenous expressed protein to the promoter of TRAF3IP2. M–O The mRNA and protein levels of TRAF3IP2 were detected in UOK109 cells and 786-O cells transfected dCas9-SETDB1 and gTRAF3IP2. P Schematic diagram for the mechanisms of that TRAF3IP2 and NONO-TFE3 fusion accelerate tumor progression of NONO-TFE3 tRCC and that lncRNA TRAF3IP2-AS1 recruits HNRNPK, DNMT1 and SETDB1 to the promoter region of TRAF3IP2 and represses the expression of TRAF3IP2 via mediating 5mC on DNA and H3K9me3. The data are presented as the mean ± SD, *P < 0.05, **P < 0.01, ***P < 0.001