Fig. 6: FAT1 loss leads to an elevated mitotic error rate and results in WGD.
a, Representative dot plots demonstrating FAT1 ablation in PC9 cells and assessment of EdU incorporation beyond the normal G2 phase, to visualize WGD. b, Top, representative western blot validating FAT1 knockout. Bottom, quantification of EdU incorporation beyond the normal G2 population showing that FAT1 knockout significantly increases the WGD population in PC9 cells. The data represent means ± s.d. Statistical significance was determined by one-way ANOVA with Bonferroni correction. Biological repeats: n = 7 (sgNT), n = 5 (sgFAT1 clone 1) and n = 4 (sgFAT1 clone 2). c, Schematic (left) and histogram (right) showing the impact of transient FAT1 knockdown in TERT RPE-1 cells on the promotion of WGD through mitotic bypass, as determined by live-cell imaging. The data represent means ± s.e.m. Statistical significance was determined by one-way ANOVA with Bonferroni correction. Biological repeats: n = 3 (with aphidicolin treatment) and n = 6 (without aphidicolin treatment). d,e, Schematics (left) and histograms (right) showing that transient FAT1 siRNA knockdown in TERT RPE-1 cells increases the rates of cytokinesis failure (d) and nuclear shape deformation (e), as determined by 30× magnification live-cell microscopy imaging at 20 min intervals. The data represent means ± s.e.m. Statistical significance was determined by two-sided paired t-test, At least 200 mitotic events were tracked per condition over five biological replicates. YFP, yellow fluorescent protein.
