Extended Data Fig. 1: Low dose nocodazole activates the spindle assembly checkpoint and allows rapid release into anaphase.

a, Asynchronous hTERT-RPE1 FUCCI cells were fixed and stained for centrosomes (Pericentrin). Representative images for each cell cycle phase are shown (n = 3 independent experiments). b, Asynchronous hTERT-RPE1 p53^WT or p53^KO cells were serum starved for 40 h before fixation and staining for primary cilia (Arl13b) and DNA. The percentage of cells with cilia for both p53^WT and p53^KO cells are quantified in the accompanying bar graph (mean ± s.e.m.; n = 3 independent experiments; p > 0.99 p53^WT +serum vs. p53^KO +serum, p = 0.13 p53^WT –serum vs. p53^KO –serum, p = 5.1E⁻¹⁴ p53^WT +serum vs. p53^WT –serum, p = 1E⁻¹⁴ p53^KO +serum vs. p53^KO –serum). c, hTERT-RPE1 cells treated with 25 ng/ml nocodazole (Noc) for 3 h were fixed and stained for DNA, tubulin, the centromere protein CENP-C, and either tubulin or the spindle assembly checkpoint marker BubR1 (n = 3 independent experiments with similar results). d, hTERT-RPE1 cells treated with 25 ng/ml nocodazole (Noc) for 45 min. Nocodazole was then washed out with fresh growth medium, and the cells were imaged at the indicated timepoints as they exited mitosis and entered G1. The percentage of cells in each mitotic phase for the indicated timepoints was quantified (mean ± s.d.; n = 3 independent experiments). Data were analysed using one-way ANOVA test with Tukey’s multiple comparisons (b) (ns, not significant; ****, p < 0.0001). Source numerical data are available in source data.

Source data