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. 2018 Jan 16;8:841. doi: 10.1038/s41598-018-19364-4

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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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(A) Western blot analysis of expression of MDA-MB-231, HMLE-Snail-ER and HMLE-Twist-ER cells transduced by the indicated vectors. Histone H3 was used as a loading control. (B) Phase contrast of MDA-MB-231 cells overexpressing macroH2A1.1, macroH2A1.2 or Zeb1 shRNA. Scale bar = 100 μm. (C) Flow cytometric analysis of cell-surface markers, CD44 and CD24, in the cells described in (B). (D) Relative expression of the mRNAs encoding macroH2A1.1, macroH2A1.2, E-cadherin, vimentin, and Zeb1 in the cells described in (B) as determined by real-time RT-PCR. Normalized to GAPDH mRNA to account for variability in template loading. (E) Flow cytometric analysis of cell-surface markers, CD44 and CD24, in fully mesenchymal HMLE-Snail-ER or HMLE-Twist-ER cells overexpressing macroH2A1.1, macroH2A1.2 or the control vector. (F) Relative expression of the mRNAs encoding macroH2A1.1, macroH2A1.2, E-cadherin, vimentin, and Zeb1 in the cells described in (E) as determined by real-time RT-PCR. Normalized to GAPDH mRNA to account for variability in template loading.