Fig. 3: Segregation error probabilities correlate with nuclear chromosome locations.
From: Nuclear chromosome locations dictate segregation error frequencies
a, Plot comparing aneuploidy percentages after Cpd-5 treatment as in Fig. 1c, and the percentage of LADs for every chromosome in RPE1-hTERT cells (mean + s.e.m., two-tailed Pearson's correlation coefficient, n = 2 independent DamID experiments). b,c, Representative FISH image of RPE1-hTERT cells (b) and quantification of the distance of chromosomes from the centre of the nucleus (c) (mean ± s.d., two-tailed ratio t-test, n = 99, 106, 108, 98, 106 or 107 chromosomes, respectively; scale bar, 5 μm). The experiment was performed in triplicate. d, Schematic depicting the nucleus of a cell with kinetochore pairs in different-coloured circles depending on their location relative to the two centrosomes. e,f, Representative still images (e) and quantification (f) of the alignment time of kinetochores in RPE1-hTERT CENPA-GFP/Centrin1-GFP cells (scale bar, 5 μm). Arrowheads indicate specific kinetochore pairs, circles centrosomes. The experiment was performed ten times (mean ± s.d., one-way analysis of variance with Tukey’s post hoc test, n = 38, 29 and 35 chromosome pairs, respectively). g,h, As in e,f, respectively, but instead cells were treated with 62.5 nM Cpd-5 and mis-segregations were quantified (h). Arrowheads denote kinetochore pairs (white, no error; yellow, error); scale bar, 5 μm. The experiment was performed in triplicate (mean ± s.e.m., two-tailed Fisher’s exact test, n = 43, 35 and 26 chromosomes). i–k, Representative still images (i) and quantification (j,k) of kinetochore behaviour in U2OS cells expressing CENPA-GFP/mCherry-α-tubulin (scale bar, 5 μm). Dashed lines represent the expected percentage of unaligned chromosomes (j) or errors (k), which are based on the number of polar kinetochore pairs at the start of nuclear envelope breakdown (NEBD) and the number of unaligned chromosomes at the start of metaphase. The experiment was performed 16 times (mean ± s.e.m., two-tailed Fisher’s exact test). Mis., misaligned; lag., laggard.
