FIG 11.

E5 does not substantially affect IFNK promoter methylation. (a) Diagram of the IFNK promoter showing the locations of the TATA box and five CpGs. (b) Bisulfite-sequencing analysis of the IFNK promoter in wild-type HPV16⁺ cells and E5 Stop cells. Following bisulfite modification of total DNA, the IFNK promoter was isolated by PCR and inserted into a cloning vector, and individual clones were sequenced. Each row corresponds to a separate clone, and each column corresponds to one of the five CpG sites in the promoter. Five clones each from 3 donor backgrounds were analyzed in the experiment. (c) MeDIP analysis of the CDKN2B (p15) promoter (Pro) and IFNK promoter DNA from HPV16⁺ cells and E5 Stop cells. DNA was subjected to immunoprecipitation with 0.25 μg/μl 5-methylcytosine-specific (5-hm-C) or preimmune (PI) serum. The immunoprecipitants were subjected to qPCR using primers specific for the p15 or IFNK promoter. n = 9. **, P < 0.01. The error bars indicate standard error of the mean.