Plasma S1P concentrations are elevated in obesity.

<p>Plasma S1P concentrations in chow (N = 13) and HFD (N = 14) fed mice (A); plasma S1P levels in lean (N = 6) and <i>ob/ob</i> (N = 8) mice (B) and circulating S1P in lean (N = 15) and obese (N = 10) humans (C). Data are mean ± SEM. **P&lt;0.01.</p

Plasma S1P concentrations in mice are elevated in response to fasting.

<p>Changes in body mass (A), blood glucose (B), plasma NEFA (C) and plasma S1P concentrations (D) in response to fasting. Data are mean ± SEM. *P&lt;0.05; **P&lt;0.01, N = 11–12.</p

Plasma S1P concentrations correlate with clinical indices of metabolic dysfunction in humans.

<p>Plasma S1P concentrations correlate with percent body fat (A), BMI (B), waist circumference (C), fasting plasma insulin (D), HOMA-IR (E), HbA1c (F), total (G) and LDL (H) cholesterol.</p

Differentially expressed microRNAs identified by microarray.

<p>Differentially expressed microRNAs identified by microarray.</p

Effect of miR-194 inhibition on protein expression of the oxidative phosphorylation complexes expression in L6 cells.

<p>Expression of the oxidative phosphorylation (OXPHOS) complexes I to V of the electron transport chain was measured by western blot in L6 cells 48 hours after transfection with a miR-194 inhibitor. Data are expressed as mean ± SEM (n = 3 per group). Representative blots are shown. <i>P</i>-values were determined using t- tests (*p<0.05 and **p<0.01 vs control).</p

miR-194 expression in the skeletal muscle of human participants and rat offspring and correlations with HOMA-IR.

<p>miR-194 expression was validated by qPCR in human (A) and rat samples (B) (n = 4–6 per group). Values are expressed as mean ± SEM. <i>P</i>-values were determined using 1-way ANOVA followed by Student-Newman-Keuls <i>post-hoc</i> test for human data (*p<0.05 vs healthy), or Student’s t-test for rat data (*p<0.05 vs control). Correlation between miR-194 expression and HOMA-IR in human (C) and rat (D) was assessed using Pearson’s or Spearman’s correlation test as appropriate. R and p-values are indicated on the graphs.</p

MiR-194 expression in the skeletal muscle of mice fed a high fat diet.

<p>MiR-194 expression was measured by qPCR in the skeletal muscle from mice fed a chow or a high fat diet for 8 weeks (n = 9–10 per group). Values are expressed as mean ± SEM. <i>P</i>-values were determined using Student’s t-test (*p<0.05 vs chow fed mice).</p

Phenotypic characteristics.

<p>Phenotypic characteristics.</p

Effect of miR-194 inhibition on glucose homeostasis in L6 cells.

<p>Basal or insulin-stimulated glucose uptake (A), glycogen synthesis (B) and lactate production (C) were assayed in L6 cells 48 hours after transfection with a miR-194 inhibitor. Glucose oxidation (D) was also measured in these transfected cells under basal conditions or after treatment with the mitochondrial uncoupler FCCP. Values are expressed as mean ± SEM of 4–6 individual experiments, with 4 technical replicates. <i>P</i>-values were determined using 2-way ANOVA (normally distributed data) or Kruskal-Wallis 1-way ANOVA on ranks (non-normally distributed data) followed by Student-Newman-Keuls <i>post-hoc</i> tests (* p<0.05 vs. control, # p<0.05 vs. basal).</p

Effect of miR-194 inhibition on AKT and GSK3β expression in L6 cells.

<p>Expression of the phosphorylated and total forms of AKT (A) and GSK3β (B) were measured by Western blot in L6 cells 48 hours after transfection with a miR-194 inhibitor. Ratios phosphorylated:total are expressed as mean ± SEM (n = 3 per group). Representative blots are shown below the graphs. <i>P</i>-values were determined using 2-way ANOVA (normally distributed data) or Kruskal-Wallis 1-way ANOVA on ranks (non-normally distributed data) followed by Student-Newman-Keuls <i>post-hoc</i> tests (* p<0.05 vs. control, # p<0.05 vs. basal).</p