Fatty acids activate S6K1 in vitro in the muscle cell line Sol 8.

<p>Oleic acid increased S6K1 phosphorylation in Sol 8 cells in a dose-dependent (A) and time-dependent (B) manner. Similar results were obtained with palmitoleic acid (C and D). S6K1 phosphorylation induced by palmitoleic acid was independent of branched-chain amino acids and amino acids in general (E). A.A., amino acid; B.C., branched-chain amino acid. One way ANOVA *<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001; two-tailed <i>t</i> test #<i>P</i><0.05, ##<i>P</i><0.01.</p

Brown adipose tissue thermogenesis in S6K-dko is diet independent.

<p>S6K-dko mice fed a high-fat diet have significantly lower levels of <i>Ucp-1</i> mRNA in brown adipose tissue (B) and higher sympathetic activity (D, F) compared to wt controls. When fed a fat-free diet, S6K-dko mice show no significant differences in <i>Ucp-1</i> mRNA levels in brown adipose tissue (A) but do have significantly higher sympathetic activity (C, E) compared to wt controls. BAT, brown adipose tissue; FFD, fat-free diet; HFD, high-fat diet. Two-tailed <i>t</i> test **<i>P</i><0.01, ***<i>P</i><0.001.</p

Muscle lipid beta oxidation in S6K-dko is diet regulated.

<p>S6K-dko mice fed a high-fat diet have higher muscle lipid beta oxidation, as shown by significantly higher mRNA levels of <i>Ucp-3</i> (B), <i>Cpt-1β</i> (D), <i>Accα</i> (F) and <i>Pdk-4</i> (H). No such differences were observed in mice fed a fat free diet (A, C, E, G). FFD, fat-free diet; HFD, high-fat diet. Two-tailed <i>t</i> test *<i>P</i><0.05, **<i>P</i><0.01.</p

Primer sequences for RT-PCR.

<p>Primer sequences for RT-PCR.</p

Energy expenditure levels in S6K-dko are diet regulated.

<p>Energy expenditure normalized for body weight (B, D), body surface area (F, H) or lean mass (J, L) is higher in S6K-dko than wt mice when fed a high-fat diet. No differences in energy expenditure are observed between S6K-dko and wt control mice fed a fat-free diet (A, C, E, G, I, K). BW, body weight; EE, energy expenditure; FFD, fat-free diet; HFD, high-fat diet. Two-tailed <i>t</i> test *<i>P</i><0.05, **<i>P</i><0.01 and two-way ANOVA *<i>P</i><0.05, ***<i>P</i><0.001.</p

Model of metabolism regulation by diet in absence of S6K.

<p>Mice S6K-dko show lower fat mass gain and higher glucose tolerance, energy expenditure and muscle lipid β-oxidation compared with wt mice when consuming a high fat diet. Those metabolic differences are not present in absence of dietary lipids.</p

Seven SNPs show sex difference.

a<p>Trait and sex for which the SNP was selected;</p>b<p>Gene labels state the nearest gene or the gene as published previously; details on all genes near the association signal can be found in the <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003500#pgen.1003500.s002" target="_blank">Figure S2</a>;</p>c<p>One-sided P-Values.</p>d<p>larger sample size due to one additional study that did not have hip circumference, and therefore could not contribute to WHRadjBMI.</p>e<p>smaller sample size as this SNP was not on Metabochip.</p><p>Shown are the seven SNPs with significant (at 5% false discovery rate) sex difference in the follow-up data. These seven SNPs exhibit genome-wide significant association in women (joint discovery and follow-up <i>P_women</i><5×10−8) and only two of these show nominally significant association in men (joint <i>P_men</i><0.05). The three loci MAP3K1, HSD17B4, and PPARG are shown here for the first time for their anthropometric trait association as well as for sex-difference.</p

Genome-wide scan for sex-specific genome-wide association highlights numerous loci.

<p>(a) Manhattan plot showing the men-specific (upward, up to 60,586 men) and women-specific (downward, up to 73,137 women) association P-values from the discovery with the 619 selected loci colored by the phenotype for which the locus was selected; (b) QQ-plot showing the sex-specific association P-values as observed against those expected under the null overall phenotypes (black) and for each phenotypes separately (colored).</p

Seven identified SNPs compared to previously published loci.

a<p>The Effect allele refers to a positive effect direction in the discovery stage for the trait and gender, the SNP was selected for;</p>b<p>Gene near this SNP which was published previously from sex-combined analyses.</p><p>The seven SNPs with sex difference are considered to depict a known locus, if the index SNP is close to a published top SNP (<1 cM). These include four of the previously reported sexually dimorphic WHR loci (Heid et al., Nat Genet 2010).</p

Overview of design and findings.

<p>Among the 7 identified loci, we defined those close to (<1 cM) published hits <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003500#pgen.1003500-LangoAllen1" target="_blank">[25]</a>, <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003500#pgen.1003500-Speliotes1" target="_blank">[29]</a>, <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003500#pgen.1003500-Heid1" target="_blank">[31]</a> as <i>near published hit</i>s and <i>novel</i> otherwise. Novel loci with sex-combined discovery P-value<5.8×10⁻⁵, which is the P-value cut-off corresponding to 5% FDR, were declared as loci that <i>could have been discovered also with sex-combined analysis</i>, and otherwise that these <i>would have been missed without the sex-stratified analyses</i>. FDR = false discovery rate.</p