Figure EV5. Unresolved DPCs undermine faithful mitotic chromosome segregation and cellular fitness (related to Fig 5).

1. HeLa cells transfected with indicated siRNAs were treated or not with 5‐azadC for 30 min and/or MPS1i in late S phase. Cells were then collected at the indicated times after 5‐azadC withdrawal and analysed by flow cytometry. Data are representative of three independent experiments. Proportion of cells with G2/M DNA content is indicated.
2. HeLa cells were synchronized in early S phase by double thymidine block, released and pulse‐treated 7 h later with formaldehyde for 1 h in the presence or absence of SUMOi. Following formaldehyde removal, cells were subjected to live‐cell imaging analysis, and the duration of mitosis (nuclear envelope breakdown (NEBD) to anaphase onset) was quantified (red bars, median; at least 156 cells, pooled from three independent experiments, were analysed per condition; ****P < 0.0001, ns: not significant, Mann–Whitney test).
3. Quantification of mitotic defects in cells in (B) (black bars, mean; n = 3 independent experiments; > 150 cells quantified per condition; *P < 0.05, paired t‐test).
4. Clonogenic survival of HeLa cells transfected with indicated siRNAs and subjected to indicated doses of formaldehyde for 30 min before replating (mean ± SEM; n = 2 independent experiments).