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. 2017 Jun 12;8:15751. doi: 10.1038/ncomms15751

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Copyright © 2017, The Author(s)

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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(a,b) Quantification of HDR (a) and NHEJ (b) assays in cells transfected with indicated siRNAs. The HDR (the percentage of GFP+ cells among mCherry+ cells) and NHEJ (the percentage of tdTomato+ cells among GFP+ cells) efficiencies in cells transfected with the indicated siRNAs were normalized to that in control (siCTL) cells. (c) Quantification of γH2AX foci in WT and Cdh1^−/− MEFs after 10 Gy IR. (d) Immunostaining of Rad51 in WT and Cdh1^−/− MEFs after 10 Gy IR. The cells were fixed 1 h after the irradiation, and processed for immunofluorescence analysis. (e) Survival of U2OS cells transfected with the indicated siRNAs after hydroxyurea treatment. Twenty four hours after transfection of the siRNAs, the cells were treated with the indicated concentrations of HU for 24 h and then cultured in fresh media for additional 2 days before the viability test. (f,g) Immunofluorescence analysis of BRCA1 (f) and RIF1 (g) in U2OS-Fucci cells transfected with the indicated siRNAs. Three days after the siRNA transfection, the cells were treated with 10 mM HU for 2 h and collected for the analysis. Top panel: representative images of BRCA1 and RIF1 foci in cells transfected with indicated siRNAs. Bottom panel: quantification of BRCA1 and RIF1 focus number per cell (n>218). Error bars indicate SEM. ** indicates P<0.001 (student’s t-test).