Figure 2.

Genomic DNA sequences corresponding to the Peg3- and Snrpn-imprinting control regions analyzed in this study. (A and B) CpG sites are numbered and shown in bold, and HhaI restriction sites are indicated with purple rectangles. For qPCR amplification in (A), genomic DNAs were either cut with HhaI or left untreated. Solid purple arrows represent the sequences where the primers for qPCR were designed. Dashed purple line corresponds to the sequence used for the Taqman probe. For methylation analysis in both (A) and (B), genomic DNAs were bisulfite-converted and PCR amplified using modified primers (Cs substituted by Ts) corresponding to the sequences indicated by blue and orange solid arrows, and either cloned for Sanger sequencing [see Supplementary Figure S2] or subjected to pyrosequencing. In this latter case the downstream primers were 5’-biotinylated (solid circles), and pyrosequencing was performed with the primers indicated by dashed blue arrows (modified by substituting Cs by Ts in its sequence).