Methylation profiles of WT and mutant ICRs. Each horizontal line of circles represents a unique sequence.

<p>Unmethylated and methylated CpGs are presented as open and closed circles, respectively. Multiples of the same methylation pattern are indicated to the right of each sequence. Lines above the circles indicate relative location of CTCF sites 2 and 4 (C2, C4) and the Sox-Oct motifs (SO). The mutant status and the allelic parent of origin are indicated to the upper left of each block of sequences. <b>(A)</b> Bisulfite analysis was performed on neonatal liver from three heterozygous mutant mice and <b>(B)</b> the maternally transmitted mutant ICRs from one 12 dpc embryonic liver. <b>(C)</b> Both WT and mutant bisulfite PCR products from oocytes were the result of four separate reactions each using the DNA from 10–20 oocytes. Oocytes were collected from a combination of homozygous and heterozygous mutant and WT females after superovulation. Each PCR product was cloned separately, and 3–6 clones of each were sequenced. Identical sequences from each product were counted only once. <b>(D)</b> Bisulfite analysis of maternally derived mutant alleles from heterozygous blastocysts was the result of two separate reactions of 6–10 blastocysts each. PCR products from each reaction were cloned separately.