Fig. 7: Distinct waves of intrinsic and extrinsic apoptosis following IR.
From: Homologous recombination promotes non-immunogenic mitotic cell death upon DNA damage
a, IncuCyte live imaging of a caspase-3 and caspase-7 fluorescence reporter in HeLa (representative of n = 2, mean ± standard deviation of three technical replicates). a.u., arbitrary units. b, Western blots of HeLa whole-cell extracts (representative of n = 2). c, A quantitative PCR with reverse transcription (RT–qPCR) of HeLa post 14 Gy IR (mean ± s.e.m. n = 3, repeated measures (RM) one-way ANOVA with Fisher’s LSD multiple comparisons test). The fold change is relative to the cells collected immediately before IR and normalized to 1 (red dashed line). d, All cell cycle outcomes in HeLa ± IR and/or BAX BAK DKO (mean ± s.e.m. of n = 3, except 20 Gy (n = 2); two-sided Fisher’s exact test of N). e, Multi-dimensional representation of d. The symbols represent the individual outcomes. f, Western blots of whole-cell extracts from HeLa ± siRNA, BAX BAK DKO and/or 14 Gy IR (representative of n = 2). g, Western blots of whole-cell extracts from HeLa (representative of n = 2). The mitotic cells were collected by shakeoff. h, RT–qPCR of HeLa ± siRNA and/or 14 Gy IR, with a fold change to non-irradiated control siRNA normalized to 1 (mean ± s.e.m. n = 3, RM one-way ANOVA with Fisher’s LSD multiple comparisons test; the P values are relative to the control siRNA + 14 Gy). i, Relative apoptosis, as measured in a, in 14-Gy-irradiated HeLa (mean ± s.e.m. n = 4, 3, 3, 4, 4 and 3 from left to right, mixed-effect one-way ANOVA with a Fisher’s LSD multiple comparisons test). j, The first mitosis outcome in asynchronous and 14-Gy-irradiated 3F HeLa (mean ± s.e.m. n = 3, except control siRNA (n = 4), two-sided Fisher’s exact test of N). For a–i, N = individual cells across all replicates and n = biological replicates. n.s., not significant.
