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. 2019 Jun 14;9:8632. doi: 10.1038/s41598-019-45058-6

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© The Author(s) 2019

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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mH2A isoforms differential colocalization with heterochromatin and mitotic marks in zebrafish embryos at 24 hpf stage. Transgenic zf embryos expressing mH2A1:GFP-mH2A1.1 (A,B,E) or mH2A2:GFP-mH2A2 (C,D,F) at 24 hpf stage were analysed using immunohistochemistry to detect heterochromatin markers trimethyl Histone3 lysine K27 and K9 (H3K27me3 and H3K9me3) (A–D) or mitotic nuclei Phosphohistone H3 (pHH3) (E,F). DAPI was used for nucleus labelling. Confocal microscope image projection of 24 hpf transgenic fish. (A,B) mH2A1:GFP-mH2A1.1 embryo shows YSL localization of GFP positive cells and lack of colocalization with H3K27me3 (A,B) and H3K9m3 (C) throughout the embryo body. (C,D) mH2A2:GFP-mH2A2 embryo stained with heterochromatin markers H3K27me3 (D) and H3K9me3 (E) reveals areas of colocalization. (E,F) mH2A1:GFP-mH2A1.1 (E) or mH2A2:GFP-mH2A2 (F) with labelled pHH3 mitotic nuclei shows partial colocalization of GFP-mH2A2 isoform with mitotic nuclei. Each labelling assay was conducted in triplicate with n = 40–50 embryos/immunolabeling.