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Growth medium and subconfluent cells were collected, washed in PBS and fixed in ice-cold 70% ethanol at 4°C over night. Cells were then washed twice in ice-cold PBS, stained with 20 μg/ml propidium iodide (PI, Sigma-Aldrich) in PBS, supplemented with 60 μg/ml RNAse A (Sigma-Aldrich), for 30 min at room temperature and analyzed with a BD LSR II multi-laser analytical flow cytometer (BD Biosciences). Cell cycle data was analyzed by ModFit LT 3.2 software (Verity Software House, Topsham, USA).
Copyright and License information: The Author(s). ©2016
Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
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