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. 2019 Feb 4;38(22):4232–4249. doi: 10.1038/s41388-019-0723-8

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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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Fig. 2 — Cyclin D1 augments H3K9me2. a Confocal microscopy of immunofluorescence for H3K9me2 (red) and nuclear staining with 4′,6-diamidino-2-phenylindole (DAPI; blue) in cyclin D1 wild-type and knockout mouse embryonic fibroblasts (MEFs), and cyclin D1^−/− MEFs rescued with MSCV-cyclin D1-IRES-GFP or vector control. Images demonstrate the reduction in H3K9me2 in cyclin D1^−/− cells. Scale bar, 20 μm with (b) quantitation of mean fluorescence shown as mean ± SEM. c Schematic representation of transgenic paradigm. d Immunohistochemical staining for H3K9me2 in the mammary gland of transgenic mice in which the cyclin D1 gene was deleted through Cre excision in the adult mammary glands. e The quantitation of H3K9me2 is shown as mean ± SEM for n = 10 separate mammary glands from two cyclin D1^WT (tamoxifen-treated cyclin D1^wt/wt;-Rosa26^{CreERT2/CreERT2} transgenic mice) and three cyclin D1^−/− mice (tamoxifen-treated cyclin D1^fl/fl;-Rosa26^{CreERT2/CreERT2} transgenic mice)