FIGURE 1.

RFWD3 translocates to sites of DNA damage and colocalizes with RPA. A, U2OS cells stably expressing GFP-RFWD3 were transfected with DsRed-PCNA and treated with laser microirradiation to induce DNA-damage tracts. After 30 min, cells were fixed and immunostained with γH2AX antibodies. B, GFP-RFWD3 expressing cells were irradiated as in A and monitored with live imaging microscopy. C, U2OS cells were treated (IR+) or untreated (IR−) with 10 Gy of IR. After 6 h, cells were pre-extracted with 0.1% Triton X-100, fixed and immunostained with RFWD3 and RPA antibodies. D, U2OS cells transfected with indicated siRNAs were treated with 10 Gy of IR and immunostained with indicated antibodies.