Figure 5. 1,407 significantly increases CIN in spindle assembly checkpoint (SAC)–deficient cells by altering microtubule (MT) dynamics.

(A, B, C) Frequency of mitotic abnormalities in control and Mad2^cKD RPE1 cells with and without 0.5 μM compound #1407 (A), RPE1 cells with 150 nM reversine with and without 0.5 μM compound #1407 (B), and MCF7 cells treated with 15 nM reversine and/or 0.5 μM compound #1407 (C). Data were obtained by time-lapse microscopy imaging and include at least three biological replicates. P-values are calculated from chi-squared test. (D) Quantification of time from start prophase to late metaphase for control and Mad2^cKD RPE1 cells with and without 0.5 μM compound #1407. At least 29 mitoses were analyzed per condition from a minimum of three time-lapse microscopy experiments. (E) Boxplot showing mean cell migration speed (μm/second) of RPE1 cells with or without 0.5 μM 1,407. Data include a minimum of three independent imaging experiments. P-values are calculated using a Wilcox test. (F) MT plus end growth rate in mitosis with and without 0.5 μM compound #1407. Each dot represents the average of 20 MT movements within a cell, 20 cells per condition. (G) IncuCyte-based growth curves of control and Mad2^cKD RPE1 in the presence or absence of 33 nM nocodazole at days 8–16. AUC is plotted relative to cell line controls, P-values are calculated using a Wilcoxon–Mann–Whitney test. Data for DMSO control curves are also used in Fig 4G and H. (H) Frequency of mitotic abnormalities in RPE1 cells with or without 0.5 μM compound #1407 and/or 33 nM nocodazole. Data were obtained by time-lapse microscopy imaging and include at least three biological replicates. P-values are calculated by chi-squared test. “n” referrers to the number of mitotic events per condition. “^#” refers to that the same data are also used in Fig 2E.