Extended Data Fig. 8: Replicative stress in EMT tumour cells is associated with the activation of dormant origins in a RHOJ-dependent manner that prevents micronuclei formation.

a-b, Representative images (a) and quantification (b) of micronuclei in EPCAM⁺, EPCAM⁻ and EPCAM⁻ Rhoj KO cells untreated and treated with cisplatin/5FU for 12 and 24h. Nuclei are stained with DAPI. Yellow arrowheads indicate micronuclei (a) (Scale bar, 25μm, n represents the number of nuclei pooled from five independent experiments, Kruskal-Wallis test followed by two-sided Mann-Whitney paired comparisons tests. p-values adjusted for multiple comparisons by Bonferroni correction, box boundaries represent 25th and 75th percentiles; whiskers, minimum and maximum; centre line, median). c-d, Representative images (c) and quantification (d) of fork asymmetry to measure fork stalling inEPCAM⁺, EPCAM⁻ and EPCAM⁻ Rhoj KO cells untreated and treated with cisplatin/5FU for 12 h. The degree of symmetry around the replication origin was calculated as long/short fork ratio (100 individual forks were measured for each cell line in three replicates, medians with interquartile range are shown, ANOVA with condition, experience and their interaction, p-values after two-way post-hoc Sidak tests). e, Western blot analysis of the indicated replication factors and loading control in the whole cell extract and loaded on chromatin in untreated conditions and 12h after cisplatin/5FU administration (n = 2, molecular weights (kDa) are indicated on the right side of the blots).