Fig. 8

Fig. 8

Effects of inhibiting the selective HDACs of broxbam in Huh-7 cells. (A) Activities of HDAC1, -2, -4 and -6 after treatment with bb or vs at concen-trations corresponding to their respective IC₅₀ and two-fold IC₅₀. Data are presented as the means ± SEM percentage of untreated control, which was set to 100%, from three experiments. ^*P<0.05, ^**P<0.01 vs. Ctrl. One-way ANOVA with Tukey's post hoc test was used to test for significance (B) Dose-dependent inhibitory effect of bb on the activity of HDAC6 relative to control, represented as the means ± SEM from three independent experiments. ^****P<0.0001 vs. Ctrl. One-way ANOVA with Tukey's post hoc test was used to test for significance. (C) Western blot analysis and (D) quantification of siRNA-mediated knockdown of HDAC6 protein expression in Huh7 cells transfected with control (si-mock) or HDAC6-specific (si-HDAC6) siRNAs for 48 h. Each set of si-mock and si-HDAC6 lanes represents one transfection experiment. Tubulin was used as the loading control. Analysis of (E) Ki-67 and (F) E2F3 mRNA expression by reverse transcription-quantitative PCR. The mRNA expression levels were normalised to that of 18S rRNA. Data was presented as the mean ± SEM from six independent experiments. One-way ANOVA with Tukey's post hoc test was used to test for significance. HDAC, histone deacetylase; bb, broxbam; vs, vorinostat; si, small interfering.