Figure EV2. Partial inhibition of CDC7 has a minor effect on DNA synthesis but markedly affects pS139 H2AX phosphorylation.

U2OS cells were treated with the indicated concentrations of XL413 in the presence or absence of 4 mM HU for 5 h and then labelled with 10 μM EdU 30 min prior to harvest. Samples were processed for flow cytometry analysis.

1. Mono‐parametric analysis of cell count against EdU intensity. Histograms are overlaid to appreciate changes in EdU intensity upon treatment with XL413 (blue lines) relative to the untreated control (grey lines).
2. Mono‐parametric analysis of cell count against pS139 histone H2AX intensity in HU‐treated cells. Histograms are overlaid to appreciate changes in pS139 histone H2AX intensity upon treatment (red lines) relative to untreated controls (grey lines).
3. Whole‐cell extracts were analysed by Western blotting with the indicated antibodies, and total protein stain (TPS) is used as a loading control. Reduction of phosphorylation of Ser40/41 on MCM2 is indicative of CDC7 inhibition. Data are representative of two repeat experiments.

Source data are available online for this figure.