Schematic representation of the nanoparticle and ELISA design.

<p>Peptide 5A contains biotin and carboxyfluorescein, lower case letters denote D-amino acids. Nanoparticles were synthesized by a free-radical polymerization method in a microemulsion system. N-isopropylacrylamide, N,N-dimethylacrylamide and acrylic acid were used as monomers with methylenebisacrylamide as cross-linker. Pentafluorophenyl methacrylate (PFM) was added and the nanoparticles were adorned with peptide 5A (B). An ELISA was designed to detect nanoparticles in biological fluids using the biotin and carboxyfluorescein groups present on peptide 5A (C). Streptavidin coated plates were used to capture the peptides by binding the biotin group and bound peptides were detected with a HRP conjugated mAb anti-fluorescein.</p

DGAT1 overexpression is associated with an increased mitochondrial function and biogenesis in HFD-fed rats.

<p>(<b>A</b>) Western blot analysis of PGC1α (<b>B</b>) OXPHOS complexes and (<b>C</b>) UCP3 in rat TA muscle. Data are expressed as mean ± SEM (n = 10–12). <b>^#</b> P<0.05 HFD-DGAT1 vs. HFD-control.</p

DGAT1 overexpression results in bigger lipid droplets in HFD-fed rats.

<p>(<b>A</b>) Electron microscopy revealed an increase in lipid droplet diameter in DGAT1 overexpressing TA muscle of HFD-fed rats. (<b>B</b>) Western blot analysis of OXPAT in rat TA muscle. Data are expressed as mean ± SEM (n = 10–12).</p

LC/MS chromatograms comparing different concentrations of peptide 5A in blood plasma matrix.

<p>Peptide 5A peaks in plasma matrix spiked with 0, 13.6 and 45.2 ng/mL (SRM trace: 877.7 → (412.2 + 341.1 + 946.4).</p

Three weeks of high-fat feeding is associated with an increased TA DAG content compared to rats on CHOW.

<p>DGAT1 overexpression leads to an increase in TA DAG content in rats on CHOW and tended to increase DAG content in rats on HFD. Data are expressed as mean ± SEM (n = 6). *P<0.05 CHOW-DGAT1 and HFD-control vs. CHOW-control.</p

Three weeks of high-fat feeding is not associated with an increased TA ceramide content compared to rats on CHOW.

<p>DGAT1 overexpression does not result in an increase in TA ceramide content in rats on CHOW or HFD. Data are expressed as mean ± SEM (n = 10–12).</p

Three weeks of high-fat feeding compared to rats on CHOW decreased whole-body insulin sensitivity as assessed by hyperinsulinemic eulgycemic clamp.

<p>DGAT1 overexpression in TA muscle increased muscle specific insulin sensitivity both in rats on CHOW as HFD. (<b>A</b>) Steady-state whole-body glucose infusion rates in rats on CHOW and HFD for three weeks. (<b>B</b>) 2DG uptake in TA muscle of CHOW fed rats after bolus injection of 2-deoxy-[3H]glucose under the same clamping conditions as in A. (<b>C</b>) 2DG uptake in TA muscle of HFD fed rats after bolus injection of 2-deoxy-[3H]glucose under the same clamping conditions as in A. Data are expressed as mean ± SEM (n = 10–12). *P<0.05 CHOW-DGAT1 and HFD-control vs. CHOW-control, <b>^#</b> P<0.05 HFD-DGAT1 vs. HFD-control.</p

The turnover of DAG and TAG is increased in the DGAT1 overexpressing TA muscle.

<p>(<b>A</b>) Western blotting of ATGL, (<b>B</b>) CGI58 (<b>C</b>) and ADRP in rat TA muscle. Data are expressed as mean ± SEM (n = 10–12). <b>^#</b> P<0.05 HFD-DGAT1 vs. HFD-control.</p