Fig. 5

HDAC2 participated in the regulation of Sin3A- or MeCP2-suppressed H3 acetylation on CTGF promoter in ET-1-stimulated WI-38 cells. A Schematic of the 550-bp ChIP primer located on the CTGF promoter. Cells were transfected with either Sin3A-HA (1 μg) or MeCP2-HA plasmid (1 μg) for 24 h and then stimulated with ET-1 for 20 min, which was followed by ChIP assay. Nonimmune IgG was used as a negative control. Equal amounts of the soluble cross-linked chromatin present in each PCR were checked by the input (n = 3). B Cells were transfected with Sin3A-HA plasmid (0.5 μg) or co-transfected with Sin3A-HA plasmid (0.5 μg) and HDAC2 siRNA (100 nM) for 24 h and then treated with ET-1 for 20 min. Cells were then lysed and immunoblotted with antibodies specific for histone H3, acetyl-H3, HDAC2, or HA. Bars indicate values of the mean ± SEM (n = 5). *p < 0.05 versus ET-1-treated cells, ^#p < 0.05 versus Sin3A-transfected cells with ET-1 stimulation. C Cells were transfected with MeCP2-HA plasmid (0.5 μg) or co-transfected with MeCP2-HA plasmid (0.5 μg) and HDAC2 siRNA (100 nM) for 24 h and then treated with ET-1 for 20 min. Cells were subsequently lysed and immunoblotted with antibodies specific for histone H3, acetyl-H3, HDAC2, or HA. Bars indicate values of the mean ± SEM (n = 5). *p < 0.05 versus ET-1-treated cells, ^#p < 0.05 versus MeCP2-transfected cells with ET-1 stimulation