Extended Data Fig. 10: Additional validation of the increased sensitivity of aneuploid cells to KIF18A inhibition.

a, The sensitivity of near-euploid and highly-aneuploid cancer cell lines to the knockdown of KIF18A in the DRIVE RNAi screen across multiple cell lineages. *P = 0.022; two-tailed t-test. b, The sensitivity of near-euploid and highly-aneuploid cancer cell lines to the knockdown of KIF18A in the DRIVE RNAi screen, after accounting for lineage-specific differences in gene dependency scores using linear regression. *P = 0.012; two-tailed t-test. c, The sensitivity of near-euploid and highly-aneuploid cancer cell lines to the knockdown of KIF18A in the DRIVE RNAi screen, across TP53 mutation classes. * P = 0.026; two-tailed t-test. d, The correlations between AS and the dependency on KIF18A in the DRIVE RNAi screen, for cell lines that have not undergone whole-genome duplication (that is, cell lines with basal ploidy of n = 2). Spearman’s ρ = -0.27 (P = 7e-04). e, The sensitivity of near-euploid and highly-aneuploid cancer cell lines to the knockdown of KIF18A in the DRIVE RNAi screen, after removing the effect of doubling time on gene dependency scores using linear regression. *P = 0.022; two-tailed t-test. f, The sensitivity of near-euploid and highly-aneuploid cancer cell lines without microsatellite instability (MSS lines only) to the knockdown of KIF18A in the DRIVE RNAi screen. ***P = 3e-04; two-tailed t-test. g, The sensitivity of near-euploid and highly-aneuploid cancer cell lines to the knockdown of KIF18A in the DRIVE RNAi screen, in cell lines that are WT for the 4 genes most selectively mutated in aneuploid human tumours (after TP53)¹². *P = 0.021 and P = 0.02, for CTCF and ARID1A, respectively; **P = 0.004; two-tailed t-test. h, The sensitivity of near-euploid and highly-aneuploid cancer cell lines to the knockdown of KIF18A in the DRIVE RNAi screen, after removing the effect of lineage subtype on gene dependency scores using linear regression. *P = 0.024; two-tailed t-test. i, The sensitivity of near-euploid and highly-aneuploid cancer cell lines to the knockdown of KIF18A in the DRIVE RNAi screen, after removing the effect of HET70 scores on gene dependency scores using linear regression. **P = 0.003; two-tailed t-test. j, Left: western blot of KIF18A protein expression levels in RPE1 and RPT cell lines. Right: Quantification of KIF18A expression levels (normalized to GAPDH). *P = 0.023; one-tailed t-test. Data represent the mean ± s.d.; n = 3 biological replicates. k, Relative protein expression levels of KIF18A, confirming successful KIF18A knockdown in the RPE1 and RPT cell lines 72h post-transfection. Left: western blot of KIF18A protein expression levels in RPE1, RPT1 and RPT3 before and after siRNA-mediated KIF18A knockdown. Right: Quantification of KIF18A expression levels (normalized to α-Tubulin). *P = 0.034, **P = 0.004; one-tailed t-test. Data represent the mean ± s.d.; n = 3 biological replicates. l, Time-lapse imaging-based quantification of the time from nuclear envelope breakdown (NEBD) to anaphase onset in RPE1 and RPT cell lines exposed to non-targeting or KIF18A-targeting siRNAs for 72h. ** P < 0.01; ****P < 1e-04; two-tailed t-test. m, The prevalence of micronuclei formation in HCT116 and HPT cells exposed to non-targeting or KIF18A-targeting siRNAs for 72h. n.s., P > 0.05; **P < 0.01; ***P < 0.001; two-tailed Fisher’s exact test. n, Imaging-based quantification of the prevalence of cell divisions with multipolar spindles in RPE1 and RPT cell lines treated with non-targeting control or KIF18A-targeting siRNAs for 72h. *P < 0.05; **P < 0.01; ***P < 0.001; two-tailed t-test; Error bars, s.d.; n = 3 biological replicates.