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. 2018 Sep 26;8:14375. doi: 10.1038/s41598-018-32685-8

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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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Silencing of USP45 or Spindly leads to defects in cell migration. (A) U2OS cell lines either expressing USP45 WT or KO were analysed for cytokinesis rate. The figure shows that the lack of USP45 does not affect the cytokinesis process and cells division as show the DAPI staining. Staining as follow: Actin (Phalloidin, green), nuclei (DAPI, blue). 60X magnification. Scale bars: 5 μm. (B) Images from time-lapse analysis of wound healing assay of U2OS cells treated either with siRNA control (scramble) or siRNA for Spindly (siRNA Spindly). Dotted white lines indicate the edge of the wound. Three independent experiments were performed and data in the graph represent mean ± s.d. Scale bars: 10 μm. (A,B) All experiments were performed at least three times and a representative example is shown. (C) USP45 WT or KO U2OS cells. Transient transfection of USP45 KO U2OS cells with USP45 WT construct rescued the delay defects; while instead transient transfection of KO U2OS cells with catalytic inactive USP45 (C199A) construct maintain the defect. Dotted white lines indicate the edge of the wound. Three independent experiments were performed and data in the graph represent mean ± s.d. Student t-test was used to determine the statistical significance. Scale bars: 10 μm.