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. 2019 Mar 11;9:4066. doi: 10.1038/s41598-019-39343-7

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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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Synchronized HeLa cells are analyzed for SKP2 and ISG proteins. (A) Schematics of HeLa S3 cells synchronization in G1/S arrest by double thymidine block and release into fresh medium. After the second release, samples were collected every 2 h during 12 h. (B) Cells were stained with propidium iodide for DNA content and the cell cycle profile was determined by flow cytometry. Debris and sub-G1 phase material were excluded. G0/G1 (2 N) and G2/M (4 N) phases are delineated. (C) An aliquot of cells from the samples analyzed in (B) was lysed for protein analysis. Lysates were immunoblotted, as indicated. Results are reported as the ratio of each protein to GAPDH. The ratio obtained for G1/S phase was set to 1.