Figure 4

Effect of 5-aza-dc on DNMTs, and H3K9me2. (A) Total protein from the 5-aza-dc treated and untreated cells was used to perform Western blot analysis with antibodies specific against DNMT1, DNMT2, DNMT3a and DNMT3b as described in Methods. Antibody against β-actin was also used as loading control. (B) Chromatins were extracted from cells with or without 5-aza-dc treatment as described in Methods. Chromatin immunoprecipitation was performed with antibody specific against histone H3K9me2, and recovered DNA fragments were used for Q-PCR with promoter specific primers of CASP8. Fold change of H3K9me2 occupancy was calculated based on the control (without 5-aza-dc treatment). Each bar represented the mean of three independent amplifications with standard deviation (SD).