Fig. 4: Loss of UBE2K increases total H3 levels and H3K9 trimethylation in hESCs.

a Immunoprecipitation with UBE2K and control FLAG antibodies in H9 hESCs followed by western blot with H3, H1 and UBE2K antibodies. The images are representative of three independent experiments. b Western blot analysis of H9 hESCs with antibodies to total histone H3 and H1. Relative percentage values of H3/β-actin (mean ± s.e.m., eight independent experiments) and H1/β-actin (mean ± s.e.m., three independent experiments) to NT shRNA control hESCs are presented. c qPCR analysis of histone H3 variants in H9 hESCs. Graph (relative expression to NT shRNA control hESCs) represents the mean ± s.e.m. of five independent experiments. d Western blot analysis of H9 hESCs with antibodies to H3K9me3 and total H3. Graph represents the relative percentage of H3K9me3/H3 ratio to NT shRNA control hESCs (mean ± s.e.m., eight independent experiments). e Immunocytochemistry of H9 hESCs with antibody to H3K9me3. Hoechst staining was used as a marker of nuclei. Scale bar represents 20 μm. The images are representative of three independent experiments. All the statistical comparisons were made by two-tailed Student’s t test for unpaired samples. P value: **P < 0.01, ***P < 0.001. NS not significant (P > 0.05).