Figure 3

Effect of FOXO3a on proliferation of serum-deprived HCC cells. (A) Western blot analysis of the efficiency of FOXO3a-siRNA (siRNA) knockdown compared to control cells (WT); **P<0.01, t-test. (B) Mean ± SD relative viability of HCC cells cultured in serum free media (SFM) and transfected with negative control siRNA (NC-siRNA) or FOXO3a-siRNA (a); Huh7 and Hep3B cells, SFM+NC-siRNA vs. SFM+FOXO3a-siRNA, **P < 0.01, two-way ANOVA; HepG2 cells, SFM+NC-siRNA vs. SFM+FOXO3a-siRNA, **P < 0.01, t- test. (B) Mean ± SD relative viability of HCC cells cultured in media containing 10% fetal bovine serum (FBS) and transfected with FOXO3a-siRNA (b); all P>0.05, two way ANOVA. EdU staining patterns (C) and the relative EdU-positvie cells ratio (D) of HCC cells transfected with pEGFP-c2-FOXO3a-3A plasmid or Negative control after 48h serum deprivation; **P<0.01, t-test. (E) Fluorescent images of HCC cells transfected with pEGFP-c2-FOXO3a-3A plasmid (a) and Western blot analysis of the efficiency of pEGFP-c2-FOXO3a-3A plasmid (plasmid) transfection compared to control cells (WT) (b). (F) Mean ± SD relative viability of HCC cells cultured in serum free media (SFM) transfected with pEGFP-c2-FOXO3a-3A plasmid or Negative control; compared with negative control, ***P<0.001, two-way ANOVA.