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Figure 1


Fig. 1. Endogenous hyperphosphorylated RPA32 (P-RPA) does not mark sites of DSBs in HSV-1-infected cells. Vero cells were infected with HSV-1 (A-C,G-I) or mock-infected (D-F) and fixed with 4% PFA as described in the Materials and Methods. (A-C) HSV-1 infected cells were double labeled with mouse anti-{gamma}H2AX (green) and rabbit anti-UL29 (red) to detect the nuclear localization of DSBs with respect to HSV-1 replication compartments, respectively. The merged image shown in C indicates that {gamma}H2AX clearly surround replication compartments. Mock-infected cells (D-F) and HSV-1-infected cells (G-I) were double labeled using mouse anti-{gamma}H2AX (green) and rabbit phosphospecific anti-P-RPA (red) to determine the nuclear localization of cellular DSBs and endogenous P-RPA, respectively. (J-L) Digital enlargements of an area of the infected cell nucleus show in G-I. Arrows indicate typical P-RPA foci that are exclusive of {gamma}H2AX staining. Images were obtained at 100x magnification with 2x zoom.





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