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. 2018 Dec 3;8:17536. doi: 10.1038/s41598-018-35616-9

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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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The phospho-mimetic auto-phosphorylation site mutant Chk1-DD induces G2 arrest in the absence of DNA damage. (a) 3T, 3T: Chk1-WT and 3T: Chk1-DD DT40 cell lines were treated with DOX for 16 hours and cell extracts were prepared from half of the cultures. 30 μg of each extract was then analysed by western blotting using indicated antibodies. Samples shown in the upper and lower panels were resolved on the same western blots respectively (see Supplementary Information for original images). (b) The remaining portion of each culture was fixed in ethanol, stained with antibody against phospho-serine 10 histone H3 (pH3, a marker of mitotic cells) and propidium iodide (PI), and analysed by flow cytometry for DNA content and the percentage of mitotic (pH3 positive) cells (inset values).