Fig. 4. Methylation analysis of DMRs at different imprinted loci in the absence of Lsh. The methylation status of individual DNA alleles derived from Lsh^+/+ and Lsh^–/– embryos are represented in the graph. The gene fragments were amplified for H19, Igf2r, Zac1 DMRs from sodium bisulphite-modified DNA and subjected to sequence analysis. (Top) Open boxes correspond to DMRs of each gene. The arrow indicates the transcription start site. Each row corresponds to an individual sequenced strand of DNA, and each circle represents a CpG on the strand, red circles and blue circles indicate methylated and unmethylated sites, respectively. Maternal (M) and paternal (P) alleles were distinguished by a DNA polymorphism in the DMR sequence (129/Czech: Igf2,1654A/G, U19619; Igf2r, 851C/G, L06446). ^*Parent-of-origin determined by the methylation pattern.