Fig. 3.

Stimulation with EZH2 inhibitors reduces H3K27me3 content in human pancreatic ductal epithelial cells. A Histone proteins were isolated from pancreatic ductal cells stimulated with EZH2 inhibitors and control cells using 5 M of sulfuric acid. Acid-precipitated (ppt) histone proteins were separated on Nu-Page gel followed by immunoblotting to quantify the total H3 and H2K27me3 levels using Li-CoR Odyssey. B Representative western blots of H3K27me3 and H3K27ac relative to total H3 following 2-day and 7-day stimulation with GSK126 at 10 µM, EPZ6438 at 1 µM, triptolide at 20 nM compared with vehicle control DMSO. C Quantitative analysis of H3K27ac and D H3K27me3 relative to total H3 following 2-day and 7-day stimulation with GSK126 at 10 µM, EPZ6438 at 1 µM, triptolide at 20 nM compared with vehicle control DMSO. Data are displayed as mean signal ratio of H3K27ac or H3K27me3 to total H3 ± SEM of 3 replicates with representative blots above. Each dot plot represents signal ratio of H3K27ac from one independent replicate. Each triangle plot represents signal ratio of H3K27ac or H3K27me3 from one independent replicate. Statistically significant differences were determined using Student’s t-tests against control. *P < 0.05, **P < 0.01