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. 2019 Feb 4;9:1335. doi: 10.1038/s41598-019-38579-7

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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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CDC25 inhibitors impair the execution of mitosis. (A) Phase contrast stills of HeLa cells synchronized by 2x thymidine block-release, treated with vehicle alone or UPD-787 (10 μM) 5 h upon release and visualized from 7.5 h to 21.5 h upon release (4 frames/h). (B) Western blot analysis of MPM2 epitopes and Histone H3 phosphorylation (pSer₁₀) in HeLa cells treated as in (A) and examined at 10 h upon release from the 2x thymidine block. (C) Western blot analysis of CDK1 (pThr₁₄ and Tyr₁₅, respectively) from HeLa cells treated as in (A) and examined at 10 h upon release from the 2x thymidine block. (D) Flow cytometric analysis of CDK1-pTyr₁₅ in cells treated as in (A) and examined 12 h upon release from the 2x thymidine block. Nocodazole (0.4 μg/ml) and Ro-3306 (9 μM) were used as controls for the level of CDK1 phosphorylation. Staining with secondary antibody only (left panel) was used to subtract self-fluorescence. (E) Quantification of positive events gated as shown in (D). (F) DNA content (DAPI staining) of the cells shown in D.