a, Single-cell CDK2 activity traces aligned to the time of mitosis in MCF7 cells. b, c, CDK2 activity traces aligned to the time of mitosis after siRNA knockdown of p53 or control siRNA (b) or in wild-type and p53-knockout (TP53−/−) cell lines (c). d, e, CDK2 activity traces of cells exposed to 1 h application of p53 activator (10 μM nutlin-3, 10 μM tenovin-6) 2–3 h before mitosis (marked in grey) in a wild-type cell line (d) or in a p21-knockout (CDKN1A−/−) cell line (e). f, Cumulative distribution functions of S/G2/M duration (time between geminin rising point and mitosis) as a function of the strength of a 20-min NCS pulse. MCF10A cells expressing the Fucci (geminin) reporter were pre-imaged for 13 h, then treated with the indicated concentration of NCS for 20 min and imaged for a further 48 h. Cells were selected when NCS was applied during S/G2 phase (control, n = 645 cells; 10 ng ml−1 NCS, 786 cells; 50 ng ml−1 NCS, 1,314 cells; 200 ng ml−1 NCS, 471 cells). g, Examples of CDK2 activity traces in response to a 20-min NCS pulse (200 ng ml−1) or a control pulse applied 9–11 h before mitosis in Fig. 2d.