FIG 2.

MHV68 infection leads to p53 stabilization. (A) 3T3 fibroblasts were mock infected, infected with MHV68 at an MOI of 5 PFU/cell, or treated with 1 μM doxorubicin (Doxo). Cells were treated with cycloheximide (CHX; 100 μg/ml) or vehicle 14 h postinfection or 4 h after doxorubicin treatment. Cells were harvested in RIPA buffer either immediately after CHX or vehicle treatment or every hour for the following 3 h. Proteins were resolved by SDS-PAGE, and immunoblot analyses were performed to detect p53 and β-actin as a loading control. (B) Densitometric analyses were performed on resulting immunoblots to compare levels of p53 between CHX-treated or vehicle-treated samples relative to that for the loading control β-actin, which is not affected by CHX treatment. The graph depicts the percentage of p53 remaining over time in CHX-treated cells relative to that of vehicle-treated controls at equivalent time points.