Fig 2.

BRLF1-mediated activation of lytic promoters is inhibited by CpG methylation. (A and B) HONE-1 cells were transfected with methylated or mock-treated BALF2p, BARF1p, BFLF2p, BMRF1p, BRRF1p, BGLF4p, BHLF1p, BHRF1p, and BMLF1p (A) and BLRF2p (B) pCpGL luciferase constructs in the presence or absence of BRLF1 and SG5 control vector, and luciferase assays were performed 2 days after transfection. The fold luciferase activity under each condition is shown relative to the activity of the unmethylated promoter in the presence of the control vector (set to 1). The error bars indicate +1 standard deviation calculated from 3 replicate experiments. (C) A representative immunoblot shows similar levels of cotransfected BRLF1 protein in the extracts used in the methylated (M) versus unmethylated (U) BHLF1 promoter luciferase assays; similar results were observed in other luciferase assays (data not shown).