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. 2018 Sep 21;9:3848. doi: 10.1038/s41467-018-06377-w

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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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