Fig. 2

Methylation analysis of rH19 ICR transgene by qMethyl-PCR. A Structure of the rH19 ICR transgene. The region analyzed by qMethyl-PCR (B–D) is shown beneath the enlarged rH19 ICR map. The locations of DNA methylation-sensitive BstUI sites in the rH19 ICR, as well as those in the qMethyl-PCR target sequences, are shown by verical lines. B, C Pedigree (non-transgenic individuals are not shown) and methylation level of the BstUI sites at around the CTCF site 3. Squares and circles (with their identification numbers shown above each object) represent male and female individuals, respectively. Blue and red colors indicate those inheriting the transgene paternally and maternally, respectively. Numbers in the objects represent DNA methylation levels of the transgenes determined by qMethyl-PCR (average of two technical replicates). D Methylation levels of the paternally (p, blue) or maternally (m, red) inherited transgenes were compared and mean ± SD values for each line are graphically depicted. P values < 0.05 are considered significantly different. As a control, the methylation level of the same sequence at the endogenous locus was analyzed without discriminating parental alleles (the expected value is around 50%, as shown in Fig. 3B)