Reduced colocalization between Rad51 and γH2AX foci in Pml^C62A/C65Aand Pml^−/−cells. (A) Representative images of Pml^WT, Pml^−/−, and Pml^C62A/C65A MEFs overexpressing GFP-Blm (GFP-Blm, green; DAPI, blue). Scale bar, 10 μm. (B) Representative costaining images of Blm/Pml in Pml^WT MEF overexpressing GFP-Blm (GFP-Blm, green; Pml, red; DAPI, blue). Scale bar, 10 μm. (C) Ratio of GFP-Blm foci in MEFs (n = 3). (D) Ratio of Rad51 foci in MEFs over a 24-hour time course following IR exposure (n = 3 per time point). (C-D) Two-tailed unpaired Student t test analyses were performed. *P < .05; **P < .01. (E) Representative images of irradiated MEFs immunostained for Rad51 (red), γH2AX (green), and DAPI (blue). Scale bar, 10 μm. (F) Coimmunoprecipitation experiments. Cells treated without (−) or 1 hour post-IR (+) exposure were lysed and immunoprecipitated (IP) with anti-Rad51 antibody and immunoblotted with anti-γH2AX antibody. Input is shown in the lower panels. The blots shown are representative of 3 independent experiments.