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. 2018 Apr 11;8:5811. doi: 10.1038/s41598-018-23832-2

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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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The histone variants H2AX, H2AZ, and H3.3 are predominantly present in TGC chromatin. (a,b) Protein levels of the H2A variants H2AX, H2AZ (a), and canonical H3 H3.1/3.2, and the H3 variant H3.3 (b) in TSCs and dTSCs. Upper panel shows western blotting results for H2AX, H2AZ, pan-H2A (a), H3.1/3.2, H3.3 and pan-H3 (b). Western blotting was performed using total histones equivalent to 0.5 μg of genomic DNA. Bottom panel shows H2AX, H2AZ, H3.1/3.2, and H3.3 levels in dTSCs, which were normalized to the pan-H2A (a) or pan-H3 level (b) (Fig. S1). The values were expressed as a ratio relative to the maximum average. The mean ± standard deviation (S.D.) of three independent experiments are shown. *P < 0.05 versus TSC (Student’s t-test). (c) Immunofluorescence staining of H2AX, H2AZ, H3.1/3.2, and H3.3 in TSCs and dTSCs at day 4, 6, and 8 of differentiation. Bars = 10 μm.