Fig. 2: Genomic instability is increased in monosomic cell lines.

a Representative images of cells with micronuclei and nuclei structure abnormalities. Inset shows the enlarged field (white arrowhead—nuclear abnormalities, yellow arrowhead—micronuclei). Three independent experiments were performed. Scale bar—10 µm. b Percentage of cells with micronuclei. Bar graphs display the mean ± SEM of three independent experiments. Number of cells analyzed (RPE1 P53 KO control = 1879, RM X = 2657, RM 10 = 3928, RM 10;18 = 2196, RM 13 = 2065; RPE1 P53 KD control = 2082, RM KD 13 = 2155, RM KD 7;10 = 1922, RM KD 19p = 2149). Statistical analysis was performed by comparing to the respective controls using two-tailed unpaired T-test; p-values are shown on the plot. c Quantification of time spent in mitosis. Time from nuclear envelope breakdown (NEBD) until the end of anaphase was measured. The median is shown as a black bar and whiskers are chosen to show the 1.5 of the interquartile range for four independent experiments. Number of cells analyzed (RPE1 P53 KO control = 358, RM X = 1161, RM 10 = 729, RM 10;18 = 446, RM 13 = 572; RPE1 p53 KD control = 754, RM KD 7;10 = 242, RM KD 19p = 149). Statistical analysis was performed by comparing to the respective controls using two-tailed unpaired T-test; p-value for all the comparisons was <0.0001 as shown on the plot. d. Percentage of anaphases with bridges. Bar graphs display the mean ± SEM of three independent experiments. Number of anaphases analyzed (RPE1 P53 KO control = 201, RM X = 198, RM 10 = 209, RM 10;18 = 162, RM 13 = 207, RPE1 P53 KD = 197, RM KD 13 = 144, RM KD 7;10 = 182, RM KD 19p = 189). Statistical analysis was performed by comparing to the respective controls using two-tailed unpaired T-test; p-values are shown on the plot. e Representative images shows immunofluorescence staining of the DNA damage marker γH2AX in control and RM 10;18 cell lines. DNA is stained with sytox green (green) and γH2AX (red). The figures are representative of three independent experiments. Scale bar—10 µm. f Quantification of percentage of cells with ≥3 γH2AX foci per cell. Bar graphs display the mean ± SEM of four independent experiments, except for RM 10 and RM 13 with three independent experiments. Number of cells analyzed (Control = 2148, RM X = 2217, RM 10 = 1619, RM 10;18 = 1372, RM 13 = 1029). Statistical analysis was performed by comparing to the respective controls using one-tailed unpaired T-test; p-values are shown on the plot. Source data are provided in a Source Data file.