Mitotic progression in AurAKO cells. (A) Mitotic exit. Cells were released from 1NMPP1 and stained with DAPI at the indicated time points. The cells with decondensed chromosomes were scored as interphase cells (n ≥ 52 cells for each condition, three independent experiments). (B) Time-lapse sequences of H2B-GFP–expressing cells after 1NMPP1 release (n = 28 cells for each condition). Times are given in minutes. (C) Quantitative analysis of B. The time spent in the indicated phases was determined from live-cell images. (D) Anaphase defects. γ-Tubulin (red) and DNA (blue; n = 50 cells for each condition, three independent experiments) are shown. (E–H) SAC proficiency. Taxol was added after 1NMPP1 release at the indicated concentrations (E) or at 10 nM (F and G) or 100 nM (H). Cells were collected, and mitotic index was determined at 3 h (E and F), 6 h (H), or at the indicated time points (G). In F, cells were transfected with either glyceraldehyde 3-phosphate dehydrogenase (GAPDH) siRNA or Mad2 siRNA 24 h before the experiment (n = 100 cells for each condition, three independent experiments). Bars: (B) 10 µm; (D) 5 µm. Error bars indicate means ± SD. Ctrl, control; NEBD, nuclear envelope breakdown.
