Fig. 6

In vitro effects of PB on HAMP expression in liver-derived cells. a Effect of Panobinostat 10 and 100 nM on BMP9- (0, 10, and 50 ng/ml) induced Hamp1 expression (relative to Hprt) in mouse precision cut liver slices. Data are normalized to baseline (untreated cells). N = 3 separate experiments. Two-way ANOVA by BMP9 dose and PB treatment. b Effect of Panobinostat 10 nM and BMP6 (0, 6, and 18 nM) on HAMP expression (relative to GAPDH) in HuH7 human hepatoma cells. Data are normalized to baseline for each condition. N = 3 separate experiments. Two-way ANOVA by BMP6 dose and PB treatment. c Effects of PB treatment vs DMSO on HAMP, ID1, SMAD7, and ATOH8 mRNA expression in Huh7 cells, n = 3 separate experiments (same experiment as BMP6 0 nM condition from b. d Effect on HAMP and ID1 mRNA of treatment with isoform-specific HDAC inhibitors in Huh7 cells. HDAC(s) targeted by each inhibitor presented in the text. N = 3 separate experiments. One-way ANOVA, t-test for comparison between RGFP966 and control. e Whole-cell H3K9ac in Huh7 cells treated with RGFP966 and PB. f Depiction of HAMP promoter luciferase activity from knockdown of each HDAC isoform in a previously published RNAi screen (two replicates per HDAC). g Validation of HDAC3 and HDAC10 knockdown effects on HAMP mRNA expression in Huh7 cells (n = 3 separate experiments, paired t-test). Data are means ± s.e.m. *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ 0.0001; NS, P > 0.05