Insulin sensitivity.

<p>Glucose response during an insulin tolerance test in LFD (C) and HFD (D) mice for n = 6−9 mice per group. WT (open symbols) and C5L2KO (closed symbols) were fasted for 4 hours before an ip insulin bolus was administered and blood glucose was measured over 2 hours. Data are presented as mean±SEM, two-way ANOVA analysis indicates a significant difference between WT-HFD and C5L2KO-HFD mice (p<0.05).</p

Body weight and IMCL quantification.

<p>Growth curves for LFD (A) and HFD (B) WT (open symbols) and C5L2KO (closed symbols) mice over the 10-week diet intervention, n = 6−9 mice. Body weight gain (C) for WT (white bars) and C5L2KO (black bars). Oil Red O staining quantification (D) n = 5 mice per group and representative slides (E) at 40× magnification. Laminin is stained in blue with lipid droplets in red. Data are presented as mean±SEM where *p<0.05, **p<0.01 vs. WT.</p

Mouse body weight and final plasma values.

<p>Data are presented at mean±SEM and significance was determined by Student’s T-test analysis between WT and C5L2KO on same diet, where *P<0.05 and **P<0.01. Epidydimal weight was determined in a separate cohort.</p

Skeletal muscle enzyme activity.

<p>HADH (A), CS (B) activities and HADH/CS ratio (C) for WT (white bars) and C5L2KO (black bars). Data are presented as mean±SEM for n = 6−8 mice per group, where *p<0.05 and **p<0.01 vs WT.</p

Skeletal muscle C5L2 protein content in humans.

<p>C5L2 protein content in obese and type 2 diabetic men (A). Plasma ASP (closed circles) and C5L2 protein levels (open circles) before (Pre) and after (+Exercise) 12-week training regimen in obese (B) and type 2 diabetes male participants (C). Correlation between insulin sensitivity and plasma ASP (D). Data are presented as box and whiskers (with minimum and maximum indicated) (A) was analyzed by unpaired Students t-test and mean±SEM results (B and C) were analyzed by a paired Students t-test, where **p<0.01 and ***p<0.001, #p = 0.08. Plasma ASP and insulin sensitivity were measured in 23 subjects.</p

Anthropometric measurements and plasma values for obese and type 2 diabetic men.

<p>Data are presented at mean±SEM and significance was determined by repeated measures ANOVA where “a” indicates significant subject effects and NS refers to not significant. Insulin sensitivity was measured in 16/18 obese and 15/18 type 2 diabetes participants.</p

Mitochondrial respiration rates, ROS production and OXPHOS content.

<p>Data are presented as mean±SEM and significance was determined by Student’s T-test analysis between WT and C5L2KO on same diet, where *p<0.05, #p = 0.08. State 3 (ADP-stimulated respiration) was induced upon the addition of ADP (450 uM), state 4 (leak respiration) was chemically attained with the addition of the ATP-synthase inhibitor oligomycin (1 µg/mL) and the maximally uncoupled state (maximum oxygen flux) was achieved by titrating the chemical uncoupler carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP).</p

Expression levels of the five associated genes.

<p>Rt-PCR for <i>Spata17, Gpatch2, Esrrg, Ush2a,</i> and <i>Kctd3</i> (n = 3–5). The results represent the mean ± SE. Comparing all animals (regardless of strain) with and without deposits using a t-test showed no significant difference for any of the genes.</p

Intracapillary glomerular deposits.

<p>Representative example of an unaffected NZW 6-month-old, an affected NZW 20-month-old and an unaffected C57BLKS 20-month-old male mouse using PAS (A–C), apoE (D–F), apoB (G–I), and apoA-IV (J–L) stainings. Deposits are seen in the glomerular capillary lumina (arrows). Immunohistochemistry staining shows that these deposits are strongly positive for apoE (E, arrows) compared to the younger NZW mouse (D) and a mouse of a strain negative for glomerular deposits (C57BLKS) (I). The deposits are weakly positive for apoB (H, arrows) and apoA-IV (K, arrows) compared to the 6-month old NZW mouse (G, J) and C57BLKS control (I, L).</p

Glomerular deposits express a lipid droplet surface protein.

<p>Glomeruli with intracapillary deposits show positive staining for a protein that binds to the surface of lipid droplets, perilipin-2 (B, D) in contrast to glomeruli without deposits, which are negative (A, C).</p