Methylation analysis of the Gtl2 DMR in Gtl2Δ5'NeoPat and Gtl2Δ5'Pat embryos. A) Schematic of the Gtl2 DMR region in wild type, Gtl2Δ5'Neo and Gtl2Δ5' alleles. The grey box represents the Gtl2Δ5' deletion region, the green box represents the Neo gene and the black box represents Gtl2 exon 1. The vertical arrows represent recognition sites for the Msp I/Hpa II restriction enzymes, and the loxP sites are indicated by black triangles. The methylation assay detects a 2.2-kb endogenous Hinc II fragment that spans the Gtl2 DMR. Modifications in the Gtl2Δ5'Neo and Gtl2Δ5' alleles generate 3.8-kb and 3.5-kb fragments, respectively. B) Methylation assay for the Gtl2 DMR in Gtl2Δ5'NeoPat129 and Gtl2Δ5'Pat129 embryos. C) Methylation assay for the Gtl2 DMR in Gtl2Δ5'NeoMat129 and Gtl2Δ5'Mat129 embryos. Restriction sites are, H, Hinc II, Hp, Hpa II and M, Msp I.