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. 2004 Aug;24(16):7140–7150. doi: 10.1128/MCB.24.16.7140-7150.2004

FIG. 4.

FIG. 4.

Abolition of G2/M checkpoint suppresses overreplication. HCT116 (p53+/+) cells were transfected with control siRNA oligonucleotide GL2 or geminin siRNA oligonucleotide (GEM) or cotransfected with the siRNA oligonucleotide for CHK1, CHK2, or both CHK1 and CHK2 and harvested 72 h after transfection. HCT116 (p53+/+) cells treated as in Fig. 1A for 57 h were treated with either caffeine or UCN-01 for another 15 h before harvest. The percentage of cells with overreplicated DNA was analyzed by FACS. The x axis indicates the siRNA oligonucleotides or chemical used together with either GL2 or GEM. (B) HCT116 (p53+/+) cells treated as in panel A were immunoblotted for the indicated proteins. (C) HCT116 (p53+/+) cells transfected with either GL2 or GEM were treated with either caffeine or UCN-01 for the last 15 h before harvest 72 h after transfection. The cells were then immunoblotted for the indicated proteins. (D) HCT116 (p53+/+) cells transfected with either GL2 or GEM were treated with caffeine for the last 15 h before harvest 72 h after transfection. Cells were then immunoblotted for indicated proteins or stained with propidium iodide for DNA content before flow cytometry analysis.