Fig. 4: Characterisation of mitotic consequences of GWL inhibition by C-604.

a Representative time-lapse images of aberrant mitotic events and consequential cellular defects induced by C-604. Images from one of n = 3 experiments are shown. Arrowheads indicate a cytoplasmic bridge between newborn RPE-1 cells. The asterisk indicates mitotic slippage. b Impact of C-604 on mitotic timing determined as the distance between mitotic entry and the first signs of cytokinesis or mitotic slippage. Purple points represent pooled individual measurements. White points represent medians of n = 3 independent experiments (circle, square, rhombus). Statistical significance was determined from n = 3 medians, using an unpaired two-tailed t-test. Determined p-values were adjusted for multiple testing using the Benjamini-Hochberg procedure. * p < 0.05, ** p < 0.01, *** p < 0.001. c Frequencies of defective mitotic exit. White points represent the results of n = 3 independent experiments. Bars and error bars represent means and standard deviations, respectively. (b, c) 25-50 mitotic events were analysed for each experiment, cell line and condition. d Correlation scatterplots of defective mitotic exit frequencies and magnitudes of cellular responses to C-604 treatment (Fig. 2g). Pearson correlation coefficients are indicated. e Representative image sequence demonstrating C-604-induced cellular fusions between cousin HeLa cells. An image sequence from one of three independent experiments is shown. f Cell lineages (n = 10) of HCC1395, RPE-1 and HeLa cells treated with increasing doses of C-604 for 60 h. Black and red circles represent cell death and fusion events, respectively. g, h Cell division (g) and fusion (h) events per lineage. Points represent individual lineages (n = 10 per condition). Horizontal red lines represent medians.