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

Fig. 5. Pol θ-mediated alt-NHEJ promotes the survival of NHEJ-deficient leukemic B cells after G1 DSBs.

Fig. 5

a Polq mRNA expression in untreated (NT), G1-blocked (ABLki) and released (Re1d to Re4d) Xrcc4−/− p53−/− v-Abl pro-B cells. β-actin mRNA was used as a control. Cell lines used: Xrcc4−/− p53−/− (Xr15307-3 & Xr15307-11). b Graph displaying the correlation between Polq mRNA expression (blue bar graph, left y axis) and the percentage of S-phase cells (black curve graph, right y-axis) in Xrcc4−/− p53−/− v-Abl pro-B cells. NT: untreated; ABLki: treated for 72 h with ABLki; Re1d to Re4d: released. Polq mRNA expression data represent the means of two independent Xrcc4−/− p53−/− v-Abl pro-B cell clones (Xr15307-3 and Xr15307-11) and are normalized to β-actin. c Cell counts for G1 blocked cells (ABLki) and released cells (Re - 1 to 8 days after released from ABLki treatment). Data represent the mean ± SEM. The number of independent experiments for each genotype is indicated into brackets. d Percentage of dead cells in G1 blocked (ABLKi) and released cells (Re - 1 to 5 days after released from ABLki treatment). Data represent the mean + SEM. *P < 0.05, ***P < 0.001 (Two-sided Student’s t-test). The number of independent experiments for each condition is indicated in the source data file. e 0.5 million Xrcc4−/− p53−/− v-Abl pro-B cells or Xrcc4−/− Polq−/− p53−/− v-Abl pro-B cells were injected in Rag2−/− mice. 7 days after injection, mice were treated with Dasatinib or with solvent (mock) for 21 consecutive days. f Kaplan–Meier survival analysis for cohorts of Rag2−/− animals injected with Xrcc4−/− p53−/− v-Abl pro-B cells (Xrcc4−/− p53−/−, n = 24) or Xrcc4−/− Polq−/− p53−/− v-Abl pro-B cells (Xrcc4−/− Polq−/− p53−/−, n = 17) and that were either mock-treated (black lines) or treated with Dasatinib at 10 mg/kg (red lines). Animals were monitored for 100 days. The median age of sacrifice (50% survival) after injection is shown for Xrcc4−/− p53−/− (mock, 17 days; Dasatinib, 37 days) and Xrcc4−/− Polq−/− p53−/− (mock, 27 days; Dasatinib, 88 days) genotypes. ***P = 0.000004 (Two-sided Log-rank test). Source data are provided as a Source Data file.