Body weight, body composition, and food intake of male global adult onset KO mice.

<p>A, Weekly body weight of adult onset KO (n = 15) and vehicle-treated control mice (n = 10), B, Nose-to-anus body length of control (n = 10) and adult onset KO mice (n = 12) at 20 weeks of age, C, Fat mass was increased in adult onset KO mice (n = 14) compared to controls (n = 10) in 9-week old mice (P = 0.05) and further increased in adult onset KO mice (n = 12) compared to controls (n = 10) at 20 weeks, D, Lean mass was reduced in adult onset KO mice (n = 14 and n = 12) compared to controls (n = 10 and n = 10) at both 9 weeks and 20 weeks of age, E, Food intake at 10 weeks of age during a 24-hour period in metabolic cages, control (n = 10) and adult onset KO mice (n = 14). Data are expressed as mean ± SE. In B–E, individual data points are shown. Time-course data were analysed using the repeated-measures ANOVA model, see <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003166#s4" target="_blank">Materials and Methods</a> (A). The other comparison p-values (B–E), correspond to a Welch <i>t</i>-test of the null hypothesis of no difference between genotypic groups. *P<0.05, **P<0.01, ***P<0.001.</p

–log₁₀<i>P</i> values of the 5 replicated meQTLs against genomic position, with the top SNP indexed and indicated by a diamond.

<p>Estimated recombination rates are shown in blue, and SNP LD is given by colour as shown in the legends (LD data from 1000 Genomes Nov 2010 CEU genotypes).</p

Energy expenditure and metabolism of male global adult onset KO mice.

<p>A, VO₂ consumed, B, VCO₂ produced, and C, energy expenditure (EE), adjusted for variation in lean mass using multiple linear regression (ANCOVA). D, respiratory exchange ratio (RER). Adult onset KO mice (n = 12) control mice (n = 10). Measurements were made over a 22-hour period during the dark and light phases in 18-week old mice. Data are expressed as mean ± SE and individual data points are shown. For details of the lean-mass adjustment made in panels A,B,C, see <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003166#s4" target="_blank">Materials and Methods</a>, *P<0.05, **P<0.01, ***P<0.001.</p

Genome-wide meQTL analysis. A:

<p><i>Cis</i>-meQTL quantile-quantile plot showing enrichment of association signal. Grey bands correspond to 95% confidence intervals. <b>B:</b> Significant meQTL are located close to CpG sites.</p

Flowchart showing the analysis pipeline.

<p><b>Top:</b> Association of DMH Methylation Score with phenotypes. <b>Bottom:</b> Primary <i>cis</i>-meQTL association study, followed by replication study. <b>Left:</b> Association of DMH probe sets with significant meQTLs with mRNA expression. <b>Right:</b> Text mining of meQTLs significant in the primary study.</p

Boxplots showing methylation level plotted against genotype for the 5 replicated meQTLs in both the primary study (left panels) and replication study (right panels).

<p>All SNPs passed quality control filtering and association with methylation levels in both data sets.</p

Energy expenditure and metabolism in male global germline <i>Fto</i> KO mice.

<p>A, VO₂ consumed, B, VCO₂ produced, and C, energy expenditure (EE), adjusted for variation in lean mass using multiple linear regression (ANCOVA). D, respiratory exchange ratio (RER). Measurements were made in male mice during the light and dark phases at 18-weeks of age. <i>Fto</i>^+/+ (n = 20), <i>Fto</i>^−/− (n = 9). Data are expressed as mean ± SE and individual data points are shown. For details of the lean-mass adjustment made in panels A–C, see <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003166#s4" target="_blank">Materials and Methods</a>, *P<0.05, **P<0.01, ***P<0.001.</p

Replicated meQTL hits.

<p>a) ID of probe set on DMH array,</p><p>b) chromosome.</p><p>c) genomic coordinates of probe set in build37.</p><p>d) genomic coordinates of SNP in build37.</p><p>e) coefficient of SNP effect.</p><p>f) standard error for the SNP effect.</p><p>g) p-value for the SNP effect.</p><p>h) ID of probe on Illumina 27k array,</p><p>i) genomic coordinates of CpG probed in build37.</p><p>j) one-sided p-value for the SNP effect in the direction of the original association.</p

Participant characteristics (Replication Study).

<p>Values are means ± standard deviation for each quantitative trait.</p

Body weight, body composition, and food intake in male global germline <i>Fto</i> Knockout mice.

<p>A, Male body weight WT <i>Fto</i>^+/+, (n = 10), <i>Fto</i>^−/− (n = 10), B, Body length WT <i>Fto</i>^+/+, (n = 14), <i>Fto</i>^−/− (n = 8), C, Fat mass in males at 20 weeks of age, WT <i>Fto</i>^+/+, (n = 17), <i>Fto</i>^−/− (n = 8), D, Lean mass in males at 20 weeks of age, WT <i>Fto</i>^+/+, (n = 17), <i>Fto</i>^−/− (n = 8), E, Intraperitoneal glucose tolerance test (IPGTT) area under the curve (AUC) in males at 16 weeks of age. WT <i>Fto</i>^+/+, (n = 9), <i>Fto</i>^−/− (n = 10), F, Food intake in male mice at 10 weeks of age during a 24-hour period in metabolic cages. WT <i>Fto</i>^+/+, (n = 12), <i>Fto</i>^−/− (n = 9). Data are expressed as mean ± SE and in B–F individual data points are shown. Time-course data were analysed using the repeated-measures ANOVA model (see <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003166#s4" target="_blank">Materials and Methods</a>) (A). The other comparison p-values (B–F) correspond to a Welch <i>t</i>-test of the null hypothesis of no difference between genotypic groups. *P<0.05, **P<0.01, ***P<0.001.</p