FIG 8.

p53 localizes to MHV68 replication compartments, yet viral DNA replication is not required for p53 inactivation. (A) 3T3 fibroblasts were mock infected (top) or infected with MHV68 at an MOI of 5 PFU/cell (bottom). Cells in the upper and lower panels were incubated in the presence of 20 μM BrdU for 12 h prior to fixation at 18 h postinfection. Cells were stained for indirect immunofluorescence microscopy using antibodies to the indicated proteins. (B) 3T3 fibroblasts were mock infected or infected with MHV68 at an MOI of 5 PFU/cell. Inocula were removed 1 h after adsorption, and cells were treated with cidofovir, PAA, or acyclovir as indicated. Cells were treated with vehicle (DMSO) or doxorubicin (5 μM) 4 h prior to harvest 18 h postinfection. Cells were harvested in RIPA buffer at the indicated times postinfection, and lysates were resolved by SDS-PAGE. Resolved proteins were detected by immunoblot analyses using antibodies that recognize the indicated proteins. Detection of β-actin serves as a loading control.