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Figure 1 | Molecular Cancer

Figure 1

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

Figure 1

Overexpression of CDC25B alters progression in S-phase but not replication duration. (A) U2OS cells conditionally expressing Ha-CDC25B were synchronized by double thymidine block (Materials and Methods). At the indicated time after release, Ha-CDC25B induced cells (U2OS CDC25B) or not (U2OS) were harvested and the cell cycle distribution was monitored by flow cytometry after propidium iodide staining (PI). A western blot analysis with monoclonal anti-Ha antibodies shows the Ha-CDC25B level in the induced U2OS CDC25B cells. (B) U2OS cells conditionally expressing Ha-CDC25B were synchronized in mitosis by nocodazole treatment (100 nM, 17 h). Mitotic cells were recovered by shake off at 0 h and grown in drug free medium. Expression of CDC25B was achieved by tetracycline removal at the time of release (0 h). At indicated times after CDC25B induction, cells were labeled with BrdU (30 ÎĽM, 15 min) and the percentage of BrdU positive cells was determined by flow cytometry analysis.

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