Fig. 10
Pharmacological targeting of G9a activity in CCA cells. a Western blot analysis of the distribution of G9a between nuclear chromatin faction (CF) and soluble nuclear fraction (SF) in control and KRASG12D expressing (MutKRAS) Huh28 cells. Representative blots, including C23 (nucleolin) analysis as loading control, are shown. b Effect of G9a inhibition on the distribution of G9a between CF and SF in control and MutKRAS Huh28 cells. Cells were treated with CM-272 (200 nM) for 72 h before fractionation and western blot analyses. Representative blots, including C23 (nucleolin) analysis as loading control, are shown. c Effect of CM-272 on G9a methylation status and interaction with HP1γ in control and MutKRAS Huh28 cells. Cells were treated with CM-272 (200 nM) for 72 h before immunoprecipitations with an anti-G9a antibody, an anti-pan-methyllysine antibody (Methyl-K) or with an anti-HP1γ antibody, and subsequent western blot analyses to detect G9a. Corresponding immunoprecipitation controls using normal rabbit IgG are included. Representative blots are shown. d Effect of CM-272 on G9a methylation and interaction with HP1γ in HuCCT-1 cells. Cells were treated or not with CM272 (200 nM) for 72 h before immunoprecipitations were carried out as described in c. Corresponding immunoprecipitation controls using normal rabbit IgG are included. Representative blots, including levels of HP1γ in total cell lysates are shown