Fig. 7

Ki-67 degradation in S phase causes replication and sister chromatid cohesion defects. A Scheme of the experiment. B Representative images of the experiment in (A). Cells were fixed at the end of the experiment and stained with anti-BrdU antibodies. Sample size: control = 326, auxin = 345. Scale bar 5 μm. C Quantification of the experiment in (B). The data were analysed with a chi-squared test. ns = not significant. D, H, L, P IGV tracks of the regions selected for FISH analyses showing the chromosome position, Pol2, H3K27ac, H3K4me3, CTCF, Rad21 and CBX3 ChIP seq profiles form Encode and obtained from experiments in HCT116 cells. Magenta lines at the bottom represent the location of the probe selected for the satellite of chromosome 1 (D), satellite of chromosome 15 (H), single-copy low-expression region on chromosome 1 (L) and single-copy high-expression region on chromosome 19 (P). E, I Quantification of the intensity of the FISH signals in the HCT116:Ki-67-AID and HCT116:Ki-67-AID + auxin samples for the probes in D (E) and H (I). Sample size: E control = 279, auxin = 285, and I control = 309, auxin = 274. F, J Quantification of the area occupied by the FISH signals in the HCT116:Ki-67 AID and HCT116:Ki-67 AID + auxin samples for the probes in D (F) and H (J). M, Q Ratio of the FISH signals intensity between the two sister chromatids in the HCT116:Ki-67 AID and HCT116:Ki-67 AID + auxin samples for the probes in L (M) and P (Q). Sample size: M control = 214, auxin = 205, and Q control = 214, auxin = 237. N, R Distance of FISH signals between the two sister chromatids in the HCT116:Ki-67 AID and HCT116:Ki-67 AID + auxin samples for the probes in L (N) and P (R). Sample size: N control = 2, auxin = 246, and R control = 207, auxin = 220. All the data presented in the violin plots were statistically analysed with a Wilcoxon test. ***p < 0.001. The box inside the violin represents the 75th and 25th percentile, whiskers are the upper and lower adjacent values and the line is the median. G, K, O, S Representative images of the FISH signals obtained in HCT116:Ki-67-AID and HCT116:Ki-67-AID + auxin samples for the probes in D (G), H (K), L (O) and P (S). FISH signals in magenta and DNA in blue. Scale bar 5 μm