Figure 3. Knockdown of CARM1 in HeLa cells resulted in increased chromatin association of NuRD and TIF1β and reduced histone acetylation.

(A) Efficient knockdown of CARM1 in HeLa cells using two shRNAs against CARM1 as revealed by Western blot analysis. The cells were transfected with control or shRNAs against CARM1 for 3 days. The whole cell extracts were prepared and analyzed by Western blotting using antibodies as indicated. Note knockdown of CARM1 did not affect the levels of other proteins tested. (B) Knockdown of CARM1 in HeLa cells resulted in increased chromatin association of NuRD complex and TIF1β and histone hypoacetylation. Chromatin fractions were prepared from HeLa cells treated with control or shRNAs against CARM1 for 3 days and analyzed by Western blotting using antibodies as indicated. (C) Transfection of HeLa cells by shRNA against CARM1 resulted in reduced histone acetylation as revealed by immunofluorescent staining. HeLa cells were transfected with shCARM1A as indicated and subjected to immunofluorescent staining 48 h after transfection. The shRNA transfected cells were GFP positive as the shRNA vector expressed GFP. Note the GFP positive cells were low in acetylated H3 and H4 immunostaining.