Fig. 4 | Scientific Reports

Fig. 4

From: CRISPR/Cas9 generated DSB clusters mimic complex lesions induced by high-LET radiation and shift repair from c-NHEJ to mutagenic repair pathways

Fig. 4

Strong dependence of MF on alt-EJ. (A) Western blot analysis showing successful knockout of PARP1 in A549-PARP1−/− cells. β-Actin and KAP1 signals serve as loading controls, while pKAP1-S824 is used as marker for efficient activation of the IR-induced DDR. Non-cropped images are provided in Figures S5A and S5B. (B) Fold increase in MF of A549 and A549-PARP1−/− cells, transfected with the indicated combinations of gRNAs. Data for the parental cell lines is replotted from Fig. 3E. Data represent the mean and SD from 3 experiments. Dots represent the raw values of each repeat. (C) Fold increase in MF of untreated and cells treated with PARP1 inhibitor (PJ34, PARP1i) or LIG1/3 (L67, LIG1/3i) inhibitor. Data for untreated cells is replotted from Fig. 3F. Data with PARP1 inhibitor represent the mean and SD from two biological repeats, while the data for LIG1/3 inhibition is from a single experiment. Dots represent the raw values generated in each repeat. (D) Fold increase of MF in A549-PRKDC−/− cells, transfected with the indicated gRNAs and treated with PARP1i. Data represent the mean and SD from at least two experiments. Dots indicate the raw values generated in each repeat. P-values for all panels are shown in Table S1.

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