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. 2011 Sep 6;23(1):83–90. doi: 10.1089/hum.2011.091

FIG. 3.

FIG. 3.

Methylation analysis of the promoter–transgene boundary in NTERT cells transduced with SFFV-containing vectors. NTERT keratinocytes transduced with lentiviral vectors encoding SFFV-GFP or SFFV-SP/GFP were cultured in the presence or absence of 5 μM azacitidine (aza) for a period of 40 days. DNA was extracted after 5, 12, and 40 days and treated with bisulfite before pyrosequencing and analysis. (A) Average percentages of methylation within a 341-bp boundary region of SFFV-SPINK5co including 13 CpGs for SFFV (white bars) and 8 CpGs for SPINK5co (black bars) show increasing methylation over time (*p<0.001). Methylation was significantly reduced by day 40, following azacitidine treatment compared with cells without azacitidine treatment (**p<0.001). There was no significant (ns) change within the azacitidine-treated group over time. (B) Low methylation levels within a 138-bp boundary region including 8 CpGs for SFFV (white bars) and 2 CpGs for eGFP (black bars) were detected in cells transduced with SFFV-GFP.