Effects of <i>KLF15</i> and <i>SLC25A10</i> knockdownon lipogenesis <i>in vitro</i>.

<p><b>A.</b><i>KLF15</i> and <i>SLC25A10</i> were knocked down using 40nM of siRNA in SVF-derived human adipocytes differentiated <i>in vitro</i> and expression of the genes evaluated using real-time PCR. Results were analyzed using Students t-test and are presented as relative fold change ± SD vs. negative control. <b>B.</b> SVF-derived adipocytes differentiated <i>in vitro</i> were transfected with 40 nM of siRNA against <i>KLF15</i> and <i>SLC25A10</i> for 48 hours followed by evaluation of basal and insulin-stimulated lipogenesis. Relative insulin-stimulated lipogenesis was calculated against non-targeting siRNA NegC at insulin-stimulated state. Induction of lipogenesis by insulin for NegC was minimum 3-fold in all experiments. Results are based on three to five biological/independent experiments.*p<0.05, **p<0.01 and ***p<0.001.</p

Selected pathways over-represented among differentially expressed genes between OIR and OIS women^a.

<p>Selected pathways over-represented among differentially expressed genes between OIR and OIS women<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0178485#t002fn001" target="_blank">^a</a>.</p

Genes associated with adipocyte IR also associated with systemic insulin resistance or T2D.

<p>Genes associated with adipocyte IR also associated with systemic insulin resistance or T2D.</p

Glucose uptake and insulin signaling genes differentially expressed between OIR and OIS women.

<p>Glucose uptake and insulin signaling genes differentially expressed between OIR and OIS women.</p

Clinical characteristics.

<p>Clinical characteristics.</p

Additional file 1: of The epigenetic signature of subcutaneous fat cells is linked to altered expression of genes implicated in lipid metabolism in obese women

Additional tables with data on DMS and differentially expressed genes between obese cases versus never-obese controls Table S1. Differentially methylated DNA sites linked to genes between obese and never-obese women. Table S2. Differentially expressed genes between obese and never-obese women. Table S3. Differentially DNA methylated sites accompanied by differential expression. Table S4. Differentially expressed genes with DMS in Adipogenesis, Insulin Signaling, and Fatty Acid Biosynthesis pathways. Table S5. Differentially expressed genes with DMS in candidate genes for BMI according to GWAS. Table S6. Differentially expressed genes with DMS in candidate genes for fat distribution according to GWAS. Table S7. DMS in WAT in relation to obesity. Table S8. DMS in WAT in relation to weight loss (Benton MC et al., Genome Biology 2015). Table S9. DMS in WAT in relation to BMI (Rönn T et al., Human Molecular Genetics 2015). Table S10. DMS in WAT in relation to T2D (Nilsson E et al., Diabetes 2014). (XLSX 5076 kb

RELATIONSHIP BETWEEN A SEDENTARY LIFESTYLE AND ADIPOSE INSULIN RESISTANCE

Sedentary people have insulin resistance in skeletal muscle but whether this also occurs in fat cells is unknown and was examined. Insulin inhibition of hydrolysis of triglycerides (antilipolysis) and stimulation of triglyceride formation (lipogenesis) was investigated in subcutaneous fat cells from 204 sedentary and 336 physically active subjects. Insulin responsiveness (maximum hormone effect) and sensitivity (half maximum effective concentration) were determined. In 69 women hyperinsulinemia-induced circulating fatty acid levels were measured. In 128 women adipose gene expression was analyzed. Responsiveness of insulin for antilipolysis (60% inhibition) and lipogenesis (2-fold stimulation) were similar between sedentary and active subjects. Sensitivity for both measures was about 10-fold decreased in sedentary subjects (p<0.01). However, only the association between antilipolysis sensitivity and physical activity remained significant when adjusting for body mass index, age, sex, waist-to-hip ratio, fat cell size and cardiometabolic disorders in multiple regression. Fatty acid levels decreased following hyperinsulinemia but remained higher in sedentary compared to active women (p=0.01). mRNA expression of insulin receptor and its substrates 1 and 2 was decreased in sedentary subjects. In conclusion, while the maximum effect is preserved, the sensitivity to insulin’s antilipolytic effect in subcutaneous fat cells is selectively lower in sedentary subjects. </p

Characteristics of genotyped SNPs tagging 40 of 49 known WHRadjBMI-associated loci.

<p>Characteristics of genotyped SNPs tagging 40 of 49 known WHRadjBMI-associated loci.</p

Linear regression analysis of the effect of the WHRadjBMI-increasing SNPscore on lipolysis phenotypes.

<p>Linear regression analysis of the effect of the WHRadjBMI-increasing SNPscore on lipolysis phenotypes.</p

Spearman rank correlations of obesity phenotypes with adipocyte lipolysis measures.

<p>Spearman rank correlations of obesity phenotypes with adipocyte lipolysis measures.</p