Fig. 3

Bisulfite sequencing analysis of rH19 ICR transgene. A Enlarged map of the rH19 ICR transgene. The region (gray horizontal bar) analyzed by qMethyl-PCR in Fig. 2 is shown above the enlarged map. The regions amplified by PCR for BS sequencing and the primer sets used are shown beneath the enlarged map. B Genomic DNA from tail-tip somatic cells of rat was subjected to BS sequencing. Each horizontal row represents a single DNA template molecule. Methylated and unmethylated CpG motifs are shown as filled and open circles, respectively. Positions of CTCF binding sites 1, 3 (C1 and C3) and 113-bp region are shown by solid and gray rectangles, respectively. The CpG motifs in the BstUI recognition site are marked by horizontal lines. C, D Genomic DNAs from germ cells (sperm or oocyte), blastocyst, and tail-tip somatic cells of TgM (lines 74 and 157 in C and D, respectively) inheriting the transgene either paternally (upper panels) or maternally (lower) were subjected to BS sequencing. ID numbers shown above each block correspond to those shown in Fig. 2B, C