Fig. 4.

Activity of PPM1D contributes to genomic instability in cells overexpressing cyclin E1. (A) RPE1‐cE, RPE1‐cE‐PPM1D^T1 and RPE1‐cE‐PPM1D^T2 cells were fixed after treatment with mock or doxycycline for 3 days and with EdU for last 30 min, probed with 53BP1 antibodies and imaged using Olympus ScanR. Plotted is a population of cells with more than five 53BP1 foci in G1 cells (2n, EdU−). More than 500 cells were analysed per condition (n = 3). Statistical significance was evaluated by the two‐tailed t‐test (*P < 0.05, **P < 0.01). Representative images are shown on the right, bar indicates 1 μm. (B) RPE1‐cE and RPE1‐cE‐PPM1D^T2 cells were treated with mock or doxycycline or a combination of doxycycline with PPM1Di for 3 days. Single cells were sequenced and the reads were analysed by aneufinder software. Every line represents data from a single cell (n > 30). Green indicates no difference of the focal copy number state compared to that observed in nontreated RPE1 cells, deviations are shown in red. (C) Quantification of B. Plotted is the number of focal copy number abberations per cell, bar indicates average. Statistical significance was evaluated by the Kruskal–Wallis test (*P < 0.05; ***P < 0.005; ****P < 0.0001).