FIG. 2.

HSV-1 ICP27 mutant d3-4 fails to localize ICP0 and ICP4 to the cytoplasm. (A) Protein localization. Vero cells were infected with WT HSV-1 or d3-4 at an MOI of 10. Cells were fixed at 6 or 12 hpi and processed for ICP0, ICP4, and ICP27 immunofluorescence. (B) Protein accumulation. Vero cells were mock infected or infected with WT HSV-1 or d3-4 at an MOI of 10, and total proteins were harvested at the indicated times. ICP0, ICP4, and ICP27 levels were determined by immunoblotting. As a loading control, the levels of cellular protein EEA1 were also analyzed. (C) Localization of ICP0 and ICP4 in infected HeLa cells. The experiment was conducted as for panel A, except HeLa cells were infected and fixed at 12 hpi.