Extended Data Fig. 3: RAD51 promotes mitotic arrest and mitotic death in irradiated cells.
From: Homologous recombination promotes non-immunogenic mitotic cell death upon DNA damage
A) Western blots of 3F HeLa whole cell extracts (*non-specific band, n = 1). B) HR and SSA reporters17,73 in siRNA and I-SceI transfected HeLa normalized to 1. (mean ± SEM n = 3, two-tailed paired t-test; t, df, and 95% CI in Source Data). Western blots of whole cell extracts from experimental cultures shown below (representative of n = 3). C) First mitosis duration of G1, S, and G2 irradiated 3F HeLa cultures from (D) and Fig. 2f combined (n = 3, except for control siRNA n = 5; mean ± SEM and Kruskal-Wallis uncorrected Dunn’s multiple comparisons test of N). D) Fist mitosis outcome in 14 Gy irradiated 3F HeLa (mean ± SEM, n = 3 except for control siRNA n = 5, and RAD51/POLθ siRNA n = 4; two-sided Fisher’s exact test of N). 14 Gy control siRNA samples for G1 and G2 irradiated cells are from Fig. 2f. E) HR activity as in (B) in siRNA transfected or RI(dl)-2 treated HeLa (mean ± SEM n = 2, 2, 1, 1, 1, 1, 3, 3 left to right). F) Experimental timing in (G-I). Mitotic outcomes were scored following media change. G-I) Time of mitotic entry (G), first mitotic outcome (H), and mitotic duration (G) for 3F HeLa treated with DMSO, 50 µM Rl(dl)-2, and 14 Gy IR (n = 1, 1, 2, 2, 3, 3 left to right in H; G, median ± interquartile range of N; H, mean ± SEM of n; two-sided Fisher’s exact test of N; I, mean ± SEM and Kruskal-Wallis uncorrected Dunn’s multiple comparisons test of N). All panels, N = individual cells across all replicates, n = biological replicates, ns = not significant. Source numerical data and unprocessed blots are available in source data.
