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. 2022 Nov 8;13:6732. doi: 10.1038/s41467-022-34612-y

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© The Author(s) 2022

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 Schematic of the DR-GFP reporter assay (top). I-SceI-DSB-induced HR efficiencies were determined in DR-U2OS cells upon treatment with control siRNA, two siRNA^IARS1, or siRNA^Rad51 (bottom). b Schematic of the EJ5-GFP reporter assay (top). The efficiency of NHEJ in EJ5-U2OS cells was measured after transfection of control, two siRNA^IARS1, or siRNA^Ligase4 (bottom). Data are represented as mean values ± SD (n = 4, independent experiments) in a and b. **P < 0.01, ***P < 0.001, ns, not significant by the analysis of two-tailed paired Student’s t-test. c Quantification of Rad51 foci in U2OS cells treated with control siRNA or two siRNA^IARS1 after exposure to 10 Gy ionizing radiation (IR) or sham irradiation. d Quantification of γH2AX in nuclei of U2OS cells treated with control siRNA or two siRNA^IARS1 after exposure to 10 Gy IR or sham irradiation. The mean values ± standard error of the mean of four (siRNA Ctrl and siRNA^IARS1#1 and #2) independent experiments are shown in c and d. *P < 0.05, **P < 0.01, ns, not significant by the analysis of two-tailed paired Student’s t-test. e Quantification of resected ssDNA at DSB sites using the ER-AsiSI system upon treatment with control siRNA, two siRNA^IARS1 or siRNA^CtIP. After 4-OHT treatment for 4 hr to induce DSBs, resected DNA was quantified by qPCR at the indicated nucleotides from AsiSI-induced DSBs. The mean values ± SD of three (siRNA Ctrl, siRNA^IARS1#1 and #2, and CtIP) independent experiments are shown. **P < 0.01, ***P < 0.001 by the analysis of two-tailed paired Student’s t-test. f SCE analysis in HeLa cells treated with control siRNA or two siRNA^IARS1. The mean values ± SD of SCE/Total metaphase chromosome (n = 35 cells) are shown. *P < 0.05, **P < 0.01 by the analysis of two-tailed paired Student’s t-test. g Clonogenic survival of HeLa cells upon treatment with control siRNA, two siRNA^IARS1, or siRNA^BRCA1 was measured after continuous exposure to olaparib for about 2 weeks. The mean values ± SD of five (siRNA Ctrl and siRNA^IARS1#1 and #2) or two (siRNA^BRCA1) independent experiments are shown. **P < 0.01, ns, not significant by the analysis of two-tailed paired Student’s t-test. a–g Source data are provided as a Source Data file.