Extended Data Fig. 6: Chromosomes behind the poles are more likely to mis-segregate.

a,b, Representative images (a) and quantification (b) of the distance from centromeric FISH probes to the centre of the nucleus in BJ-hTERT cells (scale bar, 5 μm, mean ± s.d., n > 70, 56, 68, 70 and 68 chromosomes, respectively). Data is pooled from two independent experiments. c,d,e, Distance of centromeric FISH probes to the centre of the nucleus in three cancer cell lines (mean ± s.d., two-tailed ratio t-test, n = 133, 112, 81 and 108; n = 125, 161, 134 and 153; n = 160, 151, 185, 160 chromosomes, respectively). Three independent experiments were performed. f. Schematic depicting the strategy to follow kinetochores with a similar distance to the metaphase plate (yellow circles). g,h,i, Representative stills (g) and quantification (h-i) of time to alignment for matched kinetochores based on them having the same distance to the metaphase plate in RPE1-hTERT CENPA-GFP Centrin1-GFP cells (scale bar, 5 μm). White circles mark the centrosomes. Experiment was performed 10 times (mean ± s.d., unpaired t-test, n = 21). j,k, As in e-g, but instead cells were treated with 62.5 nM Cpd-5 and mis-segregations were measured (scale bar, 5 μm). Experiment was performed in triplicate (mean ± s.e.m., Fisher’s exact test).