Fig. 3

JNK, ERK, and p38 were involved in ET-1-induced H3 acetylation through the regulation of HDAC2 phosphorylation and HDAC2 activity. A Cells were stimulated with ET-1 for 1, 3, 5, or 10 min and then lysed and immunoblotted with antibodies specific for HDAC2 or phospho-HDAC2 (S394). Bars indicate values of the mean ± SEM (n = 3). *p < 0.05 versus control group. B After ET-1 stimulation for 10 min, WI-38 cells were immunodetected with antibodies specific for phosphor-HDAC2 (purple), HDAC2 (green), and Sin3A (red); nuclei were detected with DAPI (blue). The nuclei is labeled by a white arrow. The cytosol is labeled by a white arrowhead. Bar, 50 μm. C Cells were pretreated with 10 μM of SP600125, U0126, or SB203580 or an equivalent vehicle control (dimethyl sulfoxide [DMSO]) for 30 min and then treated with ET-1 for 3 min. The cells were then lysed and immunoblotted with antibodies specific for HDAC2 or phospho-HDAC2 (S394). Bars indicate values of the mean ± SEM (n = 4). *p < 0.05 versus ET-1 stimulation. D Cells were pretreated with 10 μM SP600125, 10 μM U0126, or 10 μM SB203580 or DMSO for 30 min and then stimulated with ET-1 for 20 min. The cell lysates were immunoprecipitated with antibodies specific for HDAC2, and then HDAC activity was detected. Bars indicate values of the mean ± SEM (n = 4). *p < 0.05, versus ET-1 stimulation. E Cells were pretreated with 10 μM of SP600125, U0126, or SB203580 or DMSO for 30 min and then treated with ET-1 for 20 min. The cells were then lysed and immunoblotted with antibodies specific for histone H3 or acetyl-H3. Bars indicate values of the mean ± SEM (n = 5). *p < 0.05 versus ET-1stimulation