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. 2021 Feb 15;12(2):183. doi: 10.1038/s41419-021-03475-4

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© The Author(s) 2021

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 — A–D DR-U2OS (A and B) or NHEJ–Hela (C and D) cells were treated with siRNA, GSK3β inhibition (5 μM CHIR99021 HCl or 5 μM LY2090314), or positive control of VE-821 (5 μM, in HR assays) or NU-7441 (10 μM, in NHEJ assays) for 24 h, followed by I-SceI transfection. GFP-positive cells were analyzed by flow cytometry 48 h later. Data are expressed as mean ± SD from three independent experiments. (***p < 0.001, n.s. not significant, t test). CHIR, CHIR99021 HCl; LY, LY2090314. E Representative images of RAD51 foci in HCT-15 (parent) and their GSK3β-depleted single clone (KO1 and KO2) cells treated with 5 μM simmiparib (Upper) or HCT-15 cells following treatment with 5 μM simmiparib, GSK3i (10 μM CHIR99021 HCl or 5 μM LY2090314), or a combination for 48 h (Lower). Nuclei were stained with DAPI. Scale bar: 2 μm. Cells that contained five or more RAD51 foci/nucleus were considered as RAD51-positive cells. At least 50 cells were analyzed for each experiment and condition. All data are expressed as mean ± SD from three independent experiments. (**p < 0.01, ***p < 0.001, t test). SP, simmiparib; CHIR, CHIR99021 HCl; LY, LY2090314.