FIG. 8.

IF characterization of cellular recombination proteins with respect to the four-protein viral subassembly of transfected cells. Vero cells were transfected with expression plasmids bearing UL29, UL5, UL8, UL5,2 and oriS as described in Materials and Methods. At 16 to 24 h posttransfection, cells were fixed, permeabilized, and double labeled with antibodies to detect the localization of the HSV-1 major DNA-binding protein UL29 (green) and the indicated cellular recombination protein (red). The primary antibodies used were 367 (A), 39S (D, G, and J), 71-9A (B), rabbit anti-RAD51 (E), anti-NBS1 (H), and anti-Ku86 (K). Merged images are shown in panels C, F, I, and L. The foci observed in all four rows represent the four-protein subassembly UL29/UL5/UL8/UL52, which occurs at a limited number of sites within the nucleus of cells that are typically not in the S phase of the cell cycle. In our experience, the polyclonal UL29 antibody 367 stains diffusely within the nucleus in addition to a limited number of UL29 foci (A), whereas the monoclonal antibody 39S primarily detects UL29 within discrete foci (D, G, and J). Some variability in UL29 staining was always noted.