Methylation analysis of the CpG islands across the M. domestica IGF2 locus. Each Roman numeral corresponds to the CpG island indicated in Figure 2A. A) Bisulfite sequencing results for CpG islands II, V and VI. Filled circles represent methylated cytosines, and unfilled circles unmethylated cytosines. B) Methylation sensitive Southern analysis of CpG islands I and IV. The enzyme used to identify non-methylated CpGs are indicated at the top of the corresponding lane, the ladder sizes are shown to the left, and arrows to the right indicate the hybridizing bands for IGF2. C) Real time genomic PCR assay for CpG island III. Levels of PCR product from HpaII digested DNA, non-digested genomic DNA (control), HpaII digested 5-azacytidine treated DNA and non-digested, 5- azacytidine treated genomic DNA are shown. Error bars represent 99.9% confidence limits.