Fig. 4: Mechanistic insights of ESR2 and AOX1 in DSB repair.

a Relative expression levels of BRCA1/2 in hiPSCs with biallelic ESR2 knock-out when compared to wild type controls. Cells were either untreated (mock) or treated with cisplatin (37 nM, EC75) for 2 days. b Western blots and quantification of key DNA repair proteins associated with HR (p-BRCA1 [Ser1524], BRCA2, ATM), NHEJ (DNA-PKcs, LIG4), MMEJ (Polθ), and stress (HSP27) in wild type and ESR2 knock-out 409B2 hiPSCs, with or without cisplatin treatment (48 h, 0.2 µM). c Western blots and quantification of key DNA repair proteins in wild type and AOX1 knock-out 409B2 hiPSCs, with or without cisplatin treatment (48 h, 0.2 µM). d Heatmap summarizing (b). e Heatmap summarizing (c). f Representative immunostaining images of 53BP1 (Fluor 647) and nuclei counterstain of wild type, ESR2 knock-out, and AOX1 knock-out cells after treatment with and without 1 µM doxorubicin for 6 h. Images were equally increased for brightness and contrast. Unmodified images without magnification are shown in Supplementary Fig. 15–17. These include treatment of cisplatin and doxorubicin for 30 min and 6 h, and also contain γ-H2AX staining (Fluor 488). g Quantification of 53BP1 foci per nucleus corresponding to f and Supplementary Fig. 15–17. Dots indicate counts from different from independent replicates, and the median number of counted nuclei across all conditions is stated. Independent biological replicates were performed (n  =  4 for a, n = 3 for b–e except n = 2 for p-BRCA1 in b, n = 8 for untreated cells in g, n = 4 for treated cells in g). Error bars indicate the s.e.m. Source data are provided as a Source data file.