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. 2020 Oct 16;11:5239. doi: 10.1038/s41467-020-19060-w

Fig. 1. Alt-NHEJ rescues RAG-induced recombination in S-G2-M.

Fig. 1

a Schematic of DSB induction by RAG endonuclease in G1-arrested v-Abl pro-B cells. Kinetics of ABLki treatment and release are indicated. b Quantification of the percentage of S phase cells (PI+/Edu+ cells) in untreated conditions (NT), after exposure to ABLki for 72 h (ABLki) and one to five days after washing off ABLki (Re1d to Re5d). Lines represent means ± S.D. with a minimum of two independent B cell clones for each genotype. The number of independent experiments for each genotype and time point is indicated in the source data file. c Representative semi-quantitative nested PCR analysis of IgkV10–95-J4 coding join in untreated (NT), G1 blocked (ABLki) and released/cycling (Re) v-Abl pro-B cells of the indicated genotype. Il-2 gene PCR was used as a loading control. Two independent cell lines were used for each genotype: WT (12095 & 12096); p53−/− (15307 & 6920.1); Xrcc4−/− p53−/− (Xr15307-3 & Xr15307-11). Source data are provided as a Source Data file. The PCR gel is representative of n = 3 independent experiments. d Circos plots displaying IgkJ4-IgkVregion prey junction distribution from IgkJ4 coding end bait libraries. Junctions are represented as arcs originating from IgkJ4 breaks with a minimum of 5 reads per 1,000 bp bin for Xrcc4−/− G1 arrested libraries, 4 reads per 1000 bp bin for Xrcc4−/− p53−/− G1 arrested libraries and 10 reads per 1000 bp bin for Xrcc4−/− p53−/− released libraries. Released cells were harvested 4 to 6 days after washing off ABLki. e Quantification of IgkJ4-Vregion junctions in Xrcc4−/− and Xrcc4−/− p53−/− v-Abl pro-B cells. Values are the percentages of junctions relative to total mapped reads. Fold enrichment in Xrcc4−/− p53−/− released cells (Re) as compared to G1 blocked cells (ABLki) is indicated. Graph bars represent the pool of n = 4 independent experiments for Xrcc4−/− G1 blocked cells, n = 3 for Xrcc4−/− p53−/− G1 blocked cells and n = 6 for Xrcc4−/− p53−/− released cells with two independent cell lines for each genotype. See also Supplementary Table 4.