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Figure 3 | Molecular Cancer

Figure 3

From: Unscheduled expression of CDC25B in S-phase leads to replicative stress and DNA damage

Figure 3

Replication is required for Îł-H2AX labeling in cells expressing elevated levels of CDC25B. (A) U2OS cells were induced to overexpress Ha-CDC25B by tetracycline removal for 20 hours together with aphidicoline (2 ÎĽg/ml). The cells were incubated with BrdU (30 ÎĽM) for 15 min before harvesting at the indicated time (0, 3 h and 6 h) after release and staining with antibodies against Îł-H2AX for immunofluorescence and rat BrdU and mouse Îł-H2AX together with propidium iodide for flow cytometry analysis (bar = 10 ÎĽM). The indicated percentage corresponds to Îł-H2AX and BrdU positive cells. (B) The CDK inhibitor roscovitin (10 ÎĽM) was added at the time of CDC25B induction in U2OS cells (0 h) or 4 h hours after the beginning of induction. A 17 h after CDC25B induction, flow cytometry was achieved after staining with Îł-H2AX antibodies and propidium iodide. The indicated percentage corresponds to Îł-H2AX positive cells.

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