Relative level of expression from repressed alleles in samples heterozygous for both <i>H19</i> rs217727 and <i>IGF2</i> rs680.

<p>Graph shows increased expression from the <i>H19</i> repressed allele is not correlated (<i>P</i> = 0.09, r = 0.54) with expression from the <i>IGF2</i> repressed allele.</p

Number of informative heterozygous samples for <i>H19</i>, <i>IGF2</i> and <i>IGF2</i>R for each gestational age class.

<p>First trimester samples range from 6–12 weeks, term samples range from 37–42 weeks.</p

Levels of <i>H19</i> repressed allele expression and DNA methylation in human first trimester placentae.

<p>(A) Increased expression from the repressed <i>H19</i> allele is positively correlated (<i>P</i> = 0.0016, r = 0.61) with increased DNA methylation in region 1. (B & C) <i>H19</i> repressed allele expression is not correlated with DNA methylation in region 2 (<i>P</i> = 0.3626, r = 0.08) or region 3 (<i>P</i> = 0.4791, r = 0.04). Each point on the graph represents individual first trimester placenta samples. Methylation levels in each region represent the average methylation from 5 CpG sites in region 1 (Chr11∶2021011–2021070), 12 CpG sites in region 2 (Chr11∶2021011–2021070) and 5 CpG sites in region 3 (Chr11∶2019079–2019145). Genomic coordinates refer to reference assembly GRCh37/hg19.</p

Placental methylation levels in regions upstream and covering the <i>H19</i> transcription start site (TSS).

<p>Each genomic region where DNA methylation was measured is highlighted in yellow and numbered 1–3. DNA methylation levels for individual CpG loci are shown for first trimester (red circles, n = 13) and term (blue circles, n = 18) placentae. Distance (bp) of cytosine nucleotide from <i>H19</i> TSS is represented on x-axis. Each data point represents the mean methylation level for the gestational age class. * Indicates a significant difference in methylation levels between first trimester and term placentae at individual CpG sites. Error bars represent SEM and when not present SEM was too low to depict on the graph. The schematic representation below the graph highlights the regions between <i>H19</i> and <i>IGF2</i> where bisulfite DNA pyrosequencing was performed. Region 1 covers 5 CpG sites (Chr11∶2021011–2021070), region 2 covers 12 CpG sites (Chr11∶2021011–2021070) and region 3 covers 5 CpG sites (Chr11∶2019079–2019145). Genomic coordinates refer to reference assembly GRCh37/hg19.</p

Allelic expression ratios for <i>H19, IGF2</i> and <i>IGF2R</i> in the human placenta.

<p>The 50∶50 ratio represents equal expression from both alleles and 0∶100 ratio represents expression exclusively from one allele. Each point on the graph represents the allelic expression ratio measured in an individual placental sample. <i>H19</i> and <i>IGF2</i> samples are from first trimester and term placentae, <i>IGF2R</i> samples are all from first trimester placentae.</p

Relative levels of repressed allele expression in human first trimester and term placentae.

<p>ND = Not Determined.</p

Ratio of expression from each allele in human first trimester and term placentae measured by pyrosequencing.

<p>Each point on the graph represents the allelic expression ratio observed in an individual placental sample. (<b>A</b>) <i>H19</i> allelic expression ratio for each gestational age class. (<b>B</b>) Expression from the <i>H19</i> repressed allele is significantly higher (*<i>P</i><0.001) in first trimester placental samples. (<b>C-D</b>) <i>IGF2</i> allelic expression ratios are similar for each gestational age class (<b>C</b>) with no significant difference (<b>D</b>) between first trimester and term. First trimester samples are 6–12 weeks of gestation term samples are 37–42 weeks of gestation.</p

Relative expression from <i>IGF2R</i> alleles in the human placenta.

<p>Shaded bars for (A) <i>IGF2R</i> rs1570070 and (B) <i>IGF2R</i> rs998075 represent the proportion of expression (%) in first trimester placentae. (C) Allelic expression ratios for <i>IGF2R</i> measured two SNP loci in the same sample. These paired samples indicate both SNP loci are equivalent (paired <i>t</i>-test, <i>P</i> = 0.42) for evaluating <i>IGF2R</i> allele-specific expression.</p

Data Ghanipoor-Samami and Javadmanesh et al. for PLoS ONE 2018.xlsx

Real time qPCR data<br><br><br

Circulating INSL3 (A) and progesterone (B) concentrations in gravid (filled bars; N = 6) and non-gravid (open bars; N = 4) sheep (means ± SD).

<p>Statistical significance is shown only where P<0.05. (C) Correlation (r = 0.4; P<0.05) of INSL3 and progesterone values for samples collected on days 5 and 12 only, excluding day 17 (luteolysis).</p