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. 2018 Feb 15;9:688. doi: 10.1038/s41467-018-03107-0

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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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Fig. 5 — USP35 regulates Aurora B functions in mitosis. a HeLa cells were transfected with USP35i alone or in combination with Flag-USP35 or Flag-USP35^C450A. The cells were stained with a phospho-histone H3 (Ser10) antibody (left). The cells expressing green fluorescence (phospho-H3, phospho-histone H3) were counted and normalized to DAPI staining (right). At least 4000 cells per group were examined from three independent experiments. Scale bar = 50 μm. b HeLa cells transfected with USP35i alone or in combination with Flag-USP35 or Flag-USP35^C450A were synchronized by a treatment with 100 ng/mL NOC for 18 h. The cell lysates were immunoblotted using the indicated antibodies. Quantification of the phospho-histone H3 (p-H3) levels was done considering the amount of β-actin protein in each case. c HeLa cells transfected with CONi or USP35i were synchronized in prometaphase by treatment with 100 ng/mL NOC for 18 h and then treated with MG132 for 4 h prior to harvesting. A western blot analysis was utilized to detect Aurora B and phospho-histone H3 (Ser10) protein levels. The data in part a represent the mean ± SD (*P < 0.05, **P < 0.005, t-test)