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. 2004 Feb;78(3):1181–1194. doi: 10.1128/JVI.78.3.1181-1194.2004

FIG. 7.

FIG. 7.

Kinase assay to evaluate phosphorylation of IE63 as expressed in IE63 mutant viruses. IE63 was immunoprecipitated with polyclonal rabbit antiserum and phosphorylation was monitored by incubation of the immunoprecipitate with radiolabeled ATP, followed by SDS gel electrophoresis and phosphorimager detection. Panel A shows phosphorylation of IE63 in cells infected with rOka (bar 2) or rOKAORF47ΔC, an ORF47 kinase null mutant (bar 3), and uninfected melanoma cell control (bar 1). The bar graph in panel B shows the results of two independent kinase assays performed with IE63 mutant viruses and reported as a mean percentage relative to cells infected with rOKA/ORF63rev (bar 2); the other bars are uninfected melanoma cell control (bar 1), rOKA/ORF63rev[T171] (bar 3), rOKA/ORF63rev[S181] (bar 4), and rOKA/ORF63rev[S185] (bar 5). The lines indicate standard errors.