Fig. 5

SIRT7 regulates p53 mediated NOXA expression. a Western blot analysis of p53 protein levels Huh7.5 cell after SIRT7 knockdown (upper) or overexpression (lower). b Luciferase activity from Huh7.5 were transfected with luciferase reporter plasmid containing p53-binding consensus (p53-Luci) with or without p53 in the absence or presence of SIRT7 or SIRT7 H187Y (SIRT7 HY). **P < 0.01, one way ANOVA. c Huh7.5 cells were transfected with Flag-p53 in the absence (CON) or presence of SIRT7 or SIRT7 HY. ChIP assay was performed with anti-Flag antibody. **P < 0.01, Student’s t-test. d and e RT-PCR analysis of mRNA levels (d) and western blot analysis of protein levels (e) in Huh7.5 cells were untransfected or transfected with p53 in the absence or presence of SIRT7 or SIRT7 HY. *P < 0.05, Student’s t-test. f Huh7.5 cells were transfected with Flag-p53 (WT), Flag-p53 K320,373R (2KR), or Flag-p53 K320,373Q (2KQ). ChIP assay was performed with anti-Flag antibody. *P < 0.05, **P < 0.01, one way ANOVA. g RT-PCR analysis of mRNA levels in cells as in F. **P < 0.01, one way ANOVA. h Huh7.5 cells were transfected with WT, 2KR or 2KQ p53 for 24 h, cell were left untreated (UT) or treated with doxorubicin for 24 h, ChIP assay was performed with anti-Flag antibody. **P < 0.01 vs WT/DOX, one way ANOVA. i and j Hep3B cells were transfected with WT, 2KR, 2KQ or 5KR (K120,372,373,381,382R) for 24 h and treated with doxorubicin for 36 h, p53 expression were evaluated by western blot (i) and cell death were evaluated by TUNEL assay (j). **P < 0.01, ***P < 0.001, one way ANOVA. Data are representative of three independent experiments. Graphs show mean ± SEM of at least three independent experiments