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. 2019 Jan 30;9:956. doi: 10.1038/s41598-018-37507-5

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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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Profiling of G9a function in the cell cycle. (a) H3K9me1 and me2 levels were determined by western blot analysis using Odyssey CLs. The means of relative fluorescence intensity to H3 are shown in the graphs. me1: N = 3, me2: N = 4, independent experiments. Error bars indicate ± SD. *p < 0.05, **p < 0.01 and ***p < 0.001 by Student’s t-test. Compared to WT, G9a KO and tFucci(SCA)2.1 showed statistically significant differences (p < 0.05) for both H3K9me1 and me2. Original images are shown in (Fig. S3). (b) G9a target gene, AI662270 mRNA level was measured by quantitative RT-PCR. The mean values show the relative expression level to Hprt. N = 4, independent experiments. Compared to WT, all other samples showed statistically significant differences (p < 0.05). (c) ChIP analysis for H3K9me2. Upstream of AI662270, Xlr5 and Wfdc15a were measured by quantitative PCR. AI662270: N = 5, Xlr5 and Wfdc15a:N = 4 independent experiments. The mean values show percentage of H3K9me2 level to input. Compared to WT, G9a KO and hGem(1/110)-G9a-mVenus for AI662270, G9a KO and tFucci(SCA)2.1 for Xlr5 and Wfdc15a showed statistically significant differences (p < 0.05).