Fig. 2
HDAC5 is essential for SOX9 nuclear localisation in TAMR cells. a Western blot analysis of SOX9 protein in cytoplasmic and nuclear fractions from DMSO or pan-HDAC inhibitor (TSA, 8 nM) treated with MCF-7 TAMR cells for 24 h. b MTT assay of growth rate of TAMR MCF-7 cells treated with DMSO or TSA (8 nM). c Immunoprecipitation of MCF-7 TAMR cells with anti-SOX9 antibody followed by immunoblotting with an antibody against SIRT1. d Western blot of SOX9 protein in cytoplasmic and nuclear fractions from MCF-7 TAMR cells treated with DMSO or SIRT1 inhibitor EX527 (30 μM). e Western blot of SOX9 protein in cytoplasmic and nuclear fractions from MCF-7 TAMR cells treated with DMSO or HDAC1/HDAC3 inhibitor MS275 (2 μM). f Western blot of SOX9 protein in cytoplasmic and nuclear fractions from MCF-7 TAMR cells treated with DMSO or HDAC4/HDAC5 inhibitor LMK-235 (12 nM). g Western blots of HDAC4 and HDAC5 proteins in TAMR cells treated with LMK-235 (12 nM). h Immunoprecipitation analysis in MCF-7 TAMR cells with anti-SOX9 antibody followed by immunoblotting with antibodies against HDAC4 and HDAC5. i HDACs 1~11 and SIRTs 1~7 mRNA expressions were analysed in parental and TAMR MCF-7 cells. j Western blot of HDAC5 protein in MCF-7 parental and TAMR cells. Data are representative of means ± SEM of three independent experiments (unpaired t-test, *p < 0.05)
