Fig. 6: PBRM1 KO cells are sensitive to mitotic perturbation.

a Clonogenic survival of RPE1 parental and PBRM1 knockouts treated with increasing doses of the CDK1 inhibitor RO-3306 relative to DMSO-only treated cells. n = 6, mean ± SEM, data were analysed using a 2way ANOVA with Dunnett’s test, **p = 0.0053, ***p = 0.0005. b Representative image from clonogenic survival assay in (a). c Experimental outline for quantifying nuclear defects induced following CDK1 inhibition by RO-3306. d % of cells with mild (left) or severe (right, also see panel f) nuclear defects after 24 h of treatment with DMSO (-) or RO-3306 (+). n = 4, mean ± SEM, data were analysed by 2way ANOVA with Dunnett’s test, ****p < 0.0001. 600-1500 nuclei were analysed per condition. e Representative images from d, showing nuclear morphology after 24 h of treatment with DMSO (-) or RO-3306 (+). Scale bar corresponds to 40 µm. f Quantification of types of severe nuclear morphology defects in parental or PBRM1 knockout cells treated as per the schematic in c. n = 4, mean ± SEM. g Clonogenic survival of RPE1 parental and PBRM1 knockouts following CCNB1 siRNA (si1 or si2) treatment, normalised to survival after treatment with a scramble (scr) siRNA. Points correspond to independent biological replicates, n = 3, mean ± SEM, data were analysed by 2way ANOVA with Dunnett’s test, ***p = 0.001 and ***p = 0.002 for KO1 and KO2, respectively. h Representative image from clonogenic survival assay in g. i Clonogenic survival of a panel of renal cell carcinoma (RCC) cell lines, which are PBRM1-proficient (blue) or -deficient due to loss-of-function mutations (red). Survival was measured after CCNB1 depletion with two independent CCNB1 siRNAs, normalised to survival after treatment with a scramble siRNA. Points correspond to independent biological replicates, n = 4 (RCC-FG2, Caki-1, RCC-4), n = 5 (786-O), or n = 8 (769-P, Caki-2). Boxes contain the 25^th to 75^th percentiles with line at median, and whiskers extend to 10th and 90th percentiles. j Western blotting for PBRM1 in RCC cell lines used in (i). α-tubulin is used as a loading control. k Representative images in PBRM1-proficient (786-O) and -deficient (RCC-FG2) cell line from the survival assay in (i). Source data are provided as a source data file.