Fig. 6
The acetyltransferase Tip60 mediates H2A-K118 acetylation. a HeLa cells were transfected with siTip60 and arrested in mitosis and mitotic cell lysates were subjected to immunoblotting. b Lysates of HeLa cells transfected with Flag-Tip60 WT or catalytic mutant of Tip60 were blotted with indicated antibodies. c Recombinant GST-H2A protein was incubated with purified GST-Tip60 in the presence or absence of Ac-CoA and NU9056, Tip60 specific inhibitor, for in vitro acetylation assay. The acetylation levels of H2A were analyzed with an anti-H2A-K118 acetylation antibody. d Recombinant GST-H2A WT or K118 mutants were incubated with purified GST-Tip60 in the presence of Ac-CoA for in vitro acetylation assay. The acetylation levels of H2A were analyzed with an anti-H2A-K118 acetylation antibody. e Mitotic HeLa cells depleted of Tip60 were stained with anti-ACA (red), anti-H2A-K118ac (green) antibodies. The percentages of cells exhibiting the arm or centromeric level of H2A-K118ac are shown. Scale bar, 5 μm. f RSF1 WT or knockout HeLa cells were transfected with control or Tip60 siRNA and stained with anti-ACA and anti-H2A-pT120 antibodies. The percentages of cells exhibiting the arm or centromeric level of H2A-pT120 are shown. Scale bar, 5 μm
