FIG. 4.
Western blot analyses of purified complexes. (A) Protein complexes were purified from HeLa (C) or HeLaAAVtCR (R) cells infected with wt HSV-1 and then analyzed by Western blotting, using antibodies against a panel of cellular and viral proteins. (B) Reverse coimmunoprecipitation of tCRep in purified complexes containing selected cellular and viral proteins. Control HeLa (C) cells and HeLaAAVtCR (R) cells were infected with wt HSV-1 for 20 h (MOI = 5 PFU/cell). Nuclear extracts from 2 × 106 infected cells were subjected to coimmunoprecipitation, using UL12, Ku70, MSH2, PARP-1, PCNA, and PHB polyclonal antibodies as indicated. The anti-HA antibody was used as a negative control. The immunoprecipitates were then immunoblotted with an anti-Rep antibody (303.9). The arrows indicate the positions of the tCRep and Rep52 proteins, and the asterisk shows a nonspecific band.