Fig. 4: Distance of chromosomes from the nuclear centre dictates mis-segregation probability.

a,b, Representative FISH images (a) and quantification (b) of the distance of FISH probes from the centre in RPE1-hTERT cells synchronised as before and treated for 4 h with monastrol (mean ± s.d., two-tailed ratio t-test, n = 89, 110, 87 and 109 chromosomes, respectively). Data were pooled from three independent experiments (scale bar, 5 μm). c, Quantification of aneuploidy numbers determined by scKaryo-seq of RPE1-hTERT cells treated as in i, followed by release in Cpd-5, shake-off and replating. Data were pooled from three independent experiments (mean ± s.e.m., two-tailed binomial test, n = 373 aneuploid cells). d–f, Still images (d) and quantification of central (cen.) versus peripheral (per.) chromosomes 1 (arrows) and 9 (arrowheads) misaligned immediately before anaphase onset (e) and subsequently mis-segregating in RPE1-hTERT cells (f) (scale bar, 5 μm). White arrows indicate properly segregating chromosomes, while the red arrow follows a mis-segregating one.The experiment was performed in septuplicate (mean ± s.d., two-tailed Fisher’s exact test). g–i, Still images (g) and quantification of the peripherality (h) of chromosome 9 in DLD1-expressing dCas9-GFP-3xFKBP mCherry-FRB-LaminB1 after 24 h of rapalog (rapa.); scale bar, 5 μm). Cells were followed live to determine mis-segregations in Cpd-5 (i). The experiment was performed in triplicate (mean ± s.e.m., two-tailed Fisher’s exact test).