Phenotypes of AurA^KO cells after mitotic exit. (A) Cell cycle profiles. Cells were synchronized as in Fig. S1 E and, after release in mitosis, were harvested at the indicated time points and colabeled with BrdU and PI. (B) Mitotic index. Mitotic cells were scored from cells in A (n ≥ 100 cells for each condition, three independent experiments). Results were combined to G2/M fraction determined from A (three independent experiments). (C) Phase succession. Time-lapse microscopy was performed for 24 h after 1NMPP1 release. Interphase and mitosis time lengths were scored (n = 20 cells for each condition). (D) Metaphase spreads 10 h after 1NMPP1 release. Chromosomes 1, 2, 3, 4, and Z are shown with extra chromosomes in red and missing chromosomes in green. Chart indicates quantification of aneuploidy (n ≥ 20 cells for each condition, three independent experiments). (E) Representative images of AurA^KO cells at 10 h after 1NMPP1 release. γ-Tubulin (red), α-tubulin (green), and DNA (blue) are shown. (F) Kinetochore/MT attachment in mitotic AurA^KO cells. HA-SNAP-CenpA–expressing cells were collected in mitosis at indicated time points after 1NMPP1 removal and cold treated before fixation. The right images show a close-up of kinetochore/MT attachment in AurA^KO. HA-SNAP-CenpA (red) and α-tubulin (green) are shown. Arrows indicate missegregated centromeres in anaphase. (G) Quantitative analysis of undissociated CenpA observed in F (n = 20 cells for each condition). Bars: (E) 10 µm; (F, left) 5 µm; (F, right) 0.5 µm. Error bars indicate means ± SD. Ctrl, control.