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Fig. 4. The ATM-Chk2 DNA damage checkpoint pathway is activated by Cdt1 overexpression in G0-, S- and G2/M-phase in Rat-1 WB4 and CB4 cells. Rat-1 WB4 cells overexpressing wild-type T7-Cdt1 (A) and CB4 cells overexpressing T7-Cdt1 Cy (B) were arrested in quiescence, released, harvested at the indicated time points, and analyzed by flow cytometry and immunoblotting. (C) Asynchronous (AS) or quiescent Rat-1 or CB4 cells were treated with BrdU for 6 hours and then double-immunostained with anti-BrdU and anti-phosphorylated ATM antibodies.