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. 2021 Dec 22;601(7892):268–273. doi: 10.1038/s41586-021-04261-0

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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. 4 — a, Schematic of the SA-GFP reporter assay. b, I-SceI-induced SSA frequency in U2OS-SA HELQ^–/– cells and WT cells treated with the indicated short interfering RNA (siRNA). n = 17 (non-targeting siRNA (siNT)/WT), n = 7 (siHELQ 1), n = 12 (siHELQ_2), n = 10 (siHELQ 3) and n = 5 (HELQ^–/–) independent experiments. Data are mean ± s.e.m. c, SSA frequency in U2OS-SA cells treated with the indicated siRNA. n = 6 independent experiments. Data are mean ± s.e.m. d, Schematic of the EJ-RFP reporter assay. e, I-SceI-induced mutagenic end-joining (mEJ-RFP) and HR (DR-GFP) frequencies in U2OS-EJDR cells treated with the indicated siRNA. n = 7 (siNT), n = 4 (siHELQ 1), n = 2 (siHELQ 2) and n = 3 (siHELQ 3) independent experiments. Data are mean ± s.e.m. f, Schematic of Cas9-mediated DSB repair assay. g, Cas9-mediated DSB repair assay showing the frequencies of NHEJ, MMEJ and SSTR repair in HELQ^–/– and WT U2OS-DR or U2OS-SA cells. n = 5 (U2OS-DR WT and DR HELQ^–/–) and n = 4 (U2OS-SA and SA HELQ^–/–) independent experiments. Data are mean ± s.e.m. h, Schematic of the RFP-SCR reporter assay. Tr GFP, truncated GFP. i, I-SceI-induced STGC frequency in U2OS-RFP-SCR cells treated with the indicated siRNA. n = 13 (siNT) n = 11 (siHELQ 1), n = 12 (siRAD52) and n = 6 (siHELQ 1/siRAD52) independent experiments. Data are mean ± s.e.m. j, LTGC frequency as shown in i. n = 13 (siNT), n = 12 (siHELQ 1), n = 11 (siRAD52) and n = 6 (siHELQ 1/siRAD52) independent experiments. Data are mean ± s.e.m. k, The ratio of LTGC/total GC from the experiments in i and j. n = 14 (siNT), n = 12 (siHELQ 1), n = 12 (siRAD52) and n = 6 (siHELQ 1/siRAD52) independent experiments. Data are mean ± s.e.m. For b, c, e, g, i–k, statistical analysis was performed using two-tailed paired t-tests as compared with siNT, unless otherwise indicated.