HSV-2 pUs3 kinase is phosphorylated in ΔUL21 infected cells.

(A) Whole cell lysates prepared at 18 hours post infection from Vero cells infected at an MOI of 3 with the viruses indicated at the top of the panels or from mock infected cells were electrophoresed through polyacrylamide gels and proteins transferred to PVDF membranes. Membranes were probed with antisera indicated on the right side of each panel. Molecular weight markers in kDa are indicated on the left side of the panels. Asterisk indicates a slower migrating species of pUs3 that was prominent in cells infected with ΔUL21. (B) Whole cell lysates prepared at 18 hours post infection from Vero cells infected at an MOI of 3 with the viruses indicated at the top of the panels or from mock infected cells were either treated with λ protein phosphatase (λ PP) or untreated, then electrophoresed through polyacrylamide gels and proteins transferred to PVDF membranes. Membranes were probed with antisera indicated on the right side of each panel. Molecular weight markers in kDa are indicated on the left side of the panels. Asterisk indicates a slower migrating species of pUs3 that was prominent in cells infected with ΔUL21 virus. (C) Whole cell lysates prepared at 18 hours post infection from Vero cells infected at an MOI of 3 with the viruses indicated at the top of the panels or from mock infected cells were either treated with λ protein phosphatase (λ PP) or untreated, then electrophoresed through a regular (lower panel) or phos-tag SDS polyacrylamide gels and proteins transferred to PVDF membranes. Membranes were probed with antisera indicated on the right side of each panel. Molecular weight markers in kDa are indicated on the left side of the panels. Asterisk indicates a slower migrating species of pUs3 that was prominent in cells infected with ΔUL21 virus.