FIGURE 7.

RAD51C is critical for the intra-S-phase check point after replication stress. A, CL-V4B vector cells and CL-V4B cells stably transfected with WTRAD51C were either untreated or treated with CPT (1.5 μm for 3 h), nocodazole (NOC) (150 ng/ml), or both, and cells were harvested after 15 h and subjected to flow cytometry. B, experimental protocol for measuring intra-S-phase checkpoint. C, BrdU⁺ cells were monitored for the CL-V4B cell transfected with vector or WT RAD51C after treatment with 3 μm CPT for 2 h and pulse-labeled with BrdU before harvesting the cells at the indicated time and processed for analysis as described under “Experimental Procedures.” Incorporation of BrdU (y axis) with respect to total DNA content (x axis) was analyzed by flow cytometry. D, BrdU⁺ incorporation for RAD51C variants was monitored after treatment with 3 μm CPT for 2 h and pulse-labeled with BrdU before harvesting the cells at the indicated time and processed for analysis as described under “Experimental Procedures.”