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. 2020 Nov 13;11:5775. doi: 10.1038/s41467-020-19502-5

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© The Author(s) 2020

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. 7 — a GFP-Ku70 recruitment to tracks in RPE1-hTERT cells treated for 5 min with the HDAC inhibitors TSA and SAHA before 800 nm multiphoton micro-irradiation (upper panel). The mean ± SEM from >60 cells from 2 independent experiments is shown (lower panel). b Colocalization by confocal microscopy of CHD7 and HDAC1 at 365 nm UV-A tracks 15 min after damage induction in U2OS cells (left panel). Quantification of co-localized foci is shown as mean ± SD from 31 cells acquired in at least 3 independent experiments (right panel). c As in b, except for 53BP1 and HDAC1 (left panel). Quantification of co-localized foci is shown as mean ± SD from 17 cells acquired in at least 3 independent experiments (right panel). d Mutational signatures (left panel), deletion sizes (middle panel) and microhomology usage (in case of deletion formation) (right panel) at repair junctions in the GC92 reporter for NHEJ. GC92 cells were transfected with the indicated siRNAs and I-SceI expression vector. Repair junctions were amplified by PCR and Sanger-sequenced. The bars represent data obtained from 3 independent experiments. Statistical significance was calculated using the two-tailed Mann–Whitney U test. Statistical significance for microhomology usage was calculated for sequences showing at least 1 bp of microhomology. The junctions of the siLuc, siCHD7-1, and siCHD7-4 samples are the same as those presented in Fig. 4f. e Western blot analysis of phosphorylated RPA32 (S4/S8) and RPA32 expression in WT and CHD7 KO-2 U2OS cells transfected with the indicated siRNAs. Cells were examined 3 h after 10 Gy of ionizing radiation. Tubulin is a loading control. Representative blots from 2 independent experiments are shown. f Quantification of plasmid integration efficiencies in WT and CHD7 KO-2 U2OS cells transfected with the indicated siRNAs. The mean ± SEM from 2 to 3 independent experiments is shown. Data were normalized to WT, which was set to 100%. Statistical significance was calculated using the two-tailed Student’s t test. g Model for how the CHD7-HDAC1/2-dependent chromatin remodeling promotes DSB repair by error-free cNHEJ. Scale bar 10 µm. Source data are provided as a Source Data file.