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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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Examination of hGeminin degron conjugated G9a-mVenus degradation. (a) Construction of the G9a expression vectors using the Fucci system. G9a was fused hGeminin(1/110) or hCdt1(1/100) and mVenus. Additionally mVenus-G9a-F-hGem(1/110) contained 3 × Flag to the C-terminus of G9a. Furthermore, mVenus-G9a-F-hGem(1/110), hGem(1/110)-G9a-mVenus, and hCdt1(1/100)-G9a-mVenus contained linker DNA coupler1 between G9a and Gem or Cdt1 (thin line). (b) Strategy of the tFucci(SCA)2.1 analysis of iMEFs transiently expressing hGeminin(1/110) fused G9a vectors. (c) FACS analysis of the expression of mCheery and AmCyan (left panels), mVenus (middle panels), and DNA contents (right panels). Black line: total cells, blue line: AmCyan (+) cells, red line: mCherry (+) cells, and green line: mVenus(+).