Additional file 5: of Differences in expression rather than methylation at placenta-specific imprinted loci is associated with intrauterine growth restriction

Pyrosequencing confirmation of the aberrant placenta-specific methylation identified using HM450k methylation arrays. (A) Violin plots show the distribution of methylation for each DMR, as well as the median (white dot), mean (red line) and the interquartile range (black rectangle) are shown. Samples with hypomethylation defined by 0.05). (B) Violin plots show the distribution of methylation for the AIM1 and RHOBTB3 DMR, separated according to gender (blue for male and red for female). (C) Example of bisulphite PCR and sub-cloning of samples identified as hypomethylated. Each circle represents a single CpG on a DNA strand: (•) methylated cytosine, (o) unmethylated cytosine. Each row corresponds to an individual cloned sequence with the genotype indicated for heterozygous SNP incorporated into the amplicon. (PDF 809 kb)