Cells were collected and boiled in Laemmli buffer for 10 minutes. Total protein concentration of each cell extract was determined by Lowry assay (DC protein assay kit, Bio-Rad, Hercules, CA). 10-20 mg of total protein per extract was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Proteins were electrotransferred (ThermoFisher Scientific, Portsmouth, NH) to PVDF membranes (Millipore, Billerica, MA) and blocked in PBS with 0.05% Tween-20 and 5% nonfat milk for one hour at room temperature. Western blots for cHV68 protein expression were incubated with 10 mg/mL monoclonal mouse anti-cHV68 gB [76] (kind gift of Dr. Phillip Stevenson), monoclonal mouse anti-cHV68 M3 at 1:50, polyclonal rabbit anti-cHV68 v-cyclin at 1:2000 (kind gifts of Dr. Herbert Virgin), and monoclonal antibody to mouse b-actin at 1:1000 (Sigma Chemical, St. Louis, MO). Blots were washed for 45 minutes in PBS with 0.05% Tween-20, then incubated with donkey antimouse or donkey anti-rabbit antibodies at 1:6000 for one hour. Blots were then washed in PBS containing 0.1% Tween-20 for 45 minutes. Proteins were visualized using an ECL Plus Western blotting detection kit (Amersham Pharmacia Biotech).