Phosphorylated H2AX and ATM levels in cells transfected with plasmids expressing VZV ORF47, ORF66, or combined ORF47 and ORF66. MeWo cells were transfected with 5ug of plasmid DNAs containing GFP, carboxyl epitope tagged VZV ORF47, ORF66, ORF47 and ORF66, HSV UL13, or EBV BGLF4 using Lipofectamine 2000. Two days after transfection, the cells were harvested and lysates were immunoblotted with antibodies to (A) γ-H2AX, H2AX, or V5 epitope tag, or (B) phosphorylated ATM, ATM, or V5 epitope tag. (C) Melanoma cells were transfected with plasmids expressing carboxyl V5 epitope-tagged VZV ORF47, ORF66 or ORF13. Two days after transfection, VZV proteins were immunoprecipitated with anti-V5 antibody, washed in protein kinase buffer, incubated with γ32P ATP, separated by electrophoresis, and autoradiography was performed. Long and short arrows indicate VZV ORF47 and ORF66 protein kinases, respectively.