Fig. 4

ZNF498 represses p53 transcriptional activity by inhibiting p53 Ser46 phosphorylation. A ZNF498 represses p53 transcriptional activity. HepG2 (p53 wild-type) and Hep3B (p53-deficient) cells were transfected with a pG13L reporter plasmid, an internal control reporter plasmid pRL-CMV, and the indicated plasmid. After 36 h of transfection, cells were harvested for dual reporter gene assays (n = 3). The expression of ZNF498 and p53 was confirmed by Western blotting. B The indicated HepG2 cell lysates were blotted with the indicated antibodies. C HepG2 cells transfected with the indicated siRNA were processed for Western blotting. D The levels of phosphorylated p53 at Ser46 and ZNF498 in liver tumor tissue from DEN-treated mice were detected and quantified. Regression analysis comparing the protein levels of ZNF498 and phosphorylated p53 at Ser46 is shown below. E Hep3B cells were cotransfected with ZNF498 and wild-type p53 or p53S46A, and p53 activity was measured via pG13L luciferase reporter gene assays (n = 3). The expression of ZNF498, p53 and phosphorylated p53 at Ser46 was confirmed by Western blotting. ns, no significance; *P < 0.05; **P < 0.01; ***P < 0.001