WDR62 is required for synchronous poleward anaphase movements. (A) Mitotic timing from nuclear breakdown (NEBD) until anaphase onset (AO) of RPE1 EB3-GFP/H2B-mCherry cells treated with siCTRL or siWDR62, shown as cumulative frequency diagram or dot plot representing each cell with bars displaying mean ± SEM; N = 3, n = 156–203 cells; ****, P < 0.0001, two-tailed paired t test. (B and C) Percentage of chromosome segregation errors from RPE1 EB3-GFP/H2B-mCherry cells, either unsynchronized (B) or after monastrol release (C). Stack bars represent normal anaphase, anaphase with lagging chromosomes, or anaphase with chromosome bridges as shown on representative images; scale bars = 5 µm; gray stripes represent a cell with a lagging chromosome that will result in micronucleus; error bars represent mean ± SEM; N = 5, n = 125–209 cells for B; N = 3, n = 209–234 cells for C; ****, P < 0.0001; *, P = 0.042 by χ² test. (D) Timing from monastrol arrest release until anaphase onset from C; dot plot represents each cell; bars represent mean ± SEM; ****, P < 0.0001, two-tailed paired t test. n = 44 cells. (E–G) Automated spindle pole and KT of RPE1 GFP-centrin1/GFP-CENPA cells treated with indicated siRNA. Shown are anaphase A speed, derived from the distance between kinetochores and centrioles (D^C-KT) over time (E); anaphase B speed, derived from the intercentrosome distance (D^C-C) over time (F); and the maximal spread of kinetochores during their movements toward the poles in anaphase A for each condition (G); N = 3, n = 20 cells; bars represent mean ± SD; *, P < 0.05, one-way ANOVA. See also Video 4, Video 5, Video 6, Video 7, and Video 8.