FIG. 2.

Analysis of Wp methylation in low-passage-number LCLs. Genomic DNA was isolated from the indicated LCLs and from the EBV-positive Burkitt's lymphoma cell line Rael. As a negative control, DNA was also isolated from the EBV-negative Burkitt's lymphoma cell line DG75. DNA was bisulfite treated, amplified with primers designed to amplify bisulfite-modified DNA, cloned, and sequenced as previously described (9, 10). The primers used were those previously described (9, 10). The genome coordinates of the CpGs in the amplified region are shown on the left side of the panel. Also shown are the positions of known regulatory regions. ○, unmethylated CpGs; •, methylated CpGs. Each numbered column shows the data obtained from an independent bisulfite PCR amplification. The vertical arrows denote those bisulfite PCR clones for which no CpG methylation was observed. The B95-A and B95-C LCLs were established by infection of peripheral blood B cells with B95.8 virus. The Ak-C1 and Ak-C2 LCLs were established by infection of peripheral blood B cells with Akata virus. The 25-3, 22-19, 17-20, and 28-8 LCLs were established by infection of peripheral blood B cells with immortalizing virus recovered from P2HR-1 clone 16 cells transfected with a plasmid containing the EBV BamHI W, Y, and H fragments, the 3′ EBNA-leader protein-encoding exons, the entire EBNA 2 coding exon, and flanking sequences, as previously described (26, 27).