Figure 3

Ago2 and Dicer are required for Rad51 recruitment to DSBs. (A) U2OS cells were transfected with the indicated siRNAs for 48 h. Then, cells were treated with IR (5 Gy) and 1 h later immunostained with the indicated antibodies. Scale bars, 20 μm. (B) Quantification of cells positive for Rad51 and γH2AX. Data from three independent experiments were used to generate the histogram. Data are represented as mean ± SEM. (C) U2OS cells were treated with the indicated siRNAs for 48 h, lysed and the lysates were subjected to immunoblotting with the indicated antibodies. (D) Wild-type or Ago2^−/− MEF cells grown on microlaser dishes were treated with 10 μM BrdU for 24 h. The cells were then subjected to microirradiation with pulsed UVA laser (λ = 365 nm), and 1 h later immunostained with Rad51 and γH2AX antibodies. Scale bars, 20 μm. See also Supplementary information, Figure S5A and S5B. ^*P < 0.005, Student's t-test.