Fig. 6: Cycling aneuploid cells display decreased karyotype aberrations and upregulate DNA repair genes in comparison with arrested aneuploid cells.

a Experimental workflow for the assessment of genome instability levels by live-cell imaging in aneuploid cycling cells expressing H2b-GFP. b Representative images of cell divisions in the different samples. For illustration purposes, images were deconvoluted by using the Huygens software using the deconvolution express function. c Quantification of mitotic errors in 3 cell division rounds in control (n = 187) and aneuploid cycling (n = 274) cells. Cells treated with the Mps1 inhibitor just before starting the time-lapse were used as a positive control for mitotic errors (n = 83). **** indicates p < 0.0001. d Volcano plot illustrating the differentially expressed pathways between cycling and arrested aneuploid cells. Specific gene sets are highlighted in color. Significance was determined with an empirical p value, calculated using 1000 permutations, using the default GSEA parameters. P values were adjusted for multiple testing using the FDR method. e Heat-map showing the z-scores of single-sample GSEA (ssGSEA) scores for DNA damage-related gene sets. Quantification of γH2AX foci (f) and 53BP1 bodies (g) in cycling (n = 150) and arrested aneuploid (n = 150) cells upon IR exposure. Only EdU negative cells were analyzed in order to exclude the contribution of S-phase cells present in the non-arrested cell samples. Association between ssGSEA score for GOBP DNA repair (h) or GOBP DNA synthesis involved in DNA repair (i) or GOBP regulation of DNA repair (j) and proliferation capacity in top vs. bottom quartiles of cancer cell lines from⁵¹. **** indicates p < 0.0001. Ctrl, control (DMSO pulsed). Aneu cycling or cyc, aneuploid cycling cells. Aneu arrested or arr, aneuploid arrested cells. M, mother division. D, daughter division. GD, grand-daughter division. Mps1i, Mps1 inhibitor. In (e), #1 and #2 refer to biological replicates. Not irrad., not irradiated. Data are means of two biological replicates, except for data in (f) and (g) (three replicates). Error bars represent SEMs. Shaded error bands in (f), (g) are shown above and below for arrested and cycling cells, respectively. Two-sided Fisher’s test was performed for data in (c). Two-tailed paired t test was performed for data in (f) and (g). Two-tailed Mann–Whitney test was performed for data in (h–j). In (c), average values for each biological replicate are shown by colored dots (each color corresponds to a different biological replicate). Source Data are provided as a Source Data file. Drawings of schemes were made by partially utilizing extracts of figures published elsewhere¹.