Search

Analysis of placenta-specific DMRs in paired samples using the HM450k methylation arrays. (A) Heatmap of pairwise correlation coefficients of for placenta-specific DMRs in samples derived from CVS vs term placenta, multiple biopsies from the same placenta and those from multiple gestations. Numbers in the coloured squares represent the Pearson’s coefficients. (B) Heatmap of Infinium probes located in placenta-specific DMRs with loci with highly concordant methylation between samples highlighted by yellow boxes. (PDF 685 kb)

Polymorphic STR marker analysis in placenta samples. Examples of electropherogram showing biallelic peaks and the lack of deletions or uniparental disomy in samples with isolated methylation defects at ubiquitous DMRs. (PDF 479 kb)

Quantification of expression levels for imprinted transcripts in placenta samples. Microfluidic-based RT-qPCR analysis of imprinted transcripts in 50 placenta samples. Results are presented as violin plots for genes with statistically difference between IUGR and controls (Student’s two-tailed t-test, p

Methylation profiling of the LIN28B DMR in dizygotic twins. Schematic representation of the LIN28B locus, indicating the CpG island incorporating the DMR. Characterization of allelic methylation for placenta samples PL215 and PL216 samples from a twin gestation by bisulphite PCR and sub-cloning. 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. Quantification of total methylation at this region was performed using pyrosequencing. Gene coordinates are from hg19 genome build. (PDF 371 kb)

Description of main clinical characteristic of the placenta cohort. (DOCX 18 kb)

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)