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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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Establishment of G9a KO iMEFs expressing G9a-mVenus that was degraded in a specific phase of the cell cycle. (a) Strategy for the establishment of G9a KO iMEF expressing hGem(1/110)-G9a-mVenus and hCdt1(1/100)-G9a-mVenus (Fig. 3a). (b) The established cell lines were stained by Hoechst 33342 and analyzed using a FACSAria SORP apparatus. Black line: total cells, blue line: AmCyan (+) cells, red line: mCherry (+) cells and green line: mVenus (+). (c) The established cell lines were analyzed for live cell imaging for fluorescent signals. The images were excerpted during the first 24 h. mVenus (upper panels), and AmCyan and mCherry (lower panels) are shown in combination using bright field microscopy. They were photographed every 30 min. (d) Fluorescence intensities are indicated in the line graphs. Blue line: AmCyan, red line: mCherry, and green line: mVenus. (e) The expression of G9a-mVenus protein was determined by western blot. mCherry and AmCyan were also determined to confirm the sorting specificity. (−) Total cells, A: AmCyan (+) sorted cells, and C: mCherry (+) sorted cells.