Er hour per mouse (kcal/h) and B; energy expenditure relative to lean physique mass (LBM). The groups are WT fed SAT HFD (n58) and PUFA HFD (n58) at the same time as Gpr120 KO mice fed SAT HFD (n57) and PUFA HFD (n57). Imply values for power expenditure over 72 h was calculated for each and every individual mouse and the graphs show imply values for the remedy groups. Statistical analysis was performed working with 1-way ANOVA followed by Student’s t-test comparing SAT HFD and PUFA HFD in every single genotype, p,0.05. doi:10.1371/journal.pone.0114942.gPLOS 1 | DOI:ten.1371/journal.pone.0114942 December 26,11 /GPR120 Will not be Essential for n-3 PUFA Effects on Energy MetabolismBoth WT and Gpr120 KO had drastically reduce fasting insulin levels on PUFA HFD than on SAT HFD. In contrast, Gpr120 KO mice, but not WT mice, had considerably decrease fasting plasma glucose levels on PUFA HFD as in comparison with SAT HFD. An insulin resistance index (glucose (mM) x insulin (ng/ml)) was calculated and it was drastically Caspase Formulation reduced in each groups of mice on PUFA HFD than in those on SAT HFD (Fig. 5A). Oral glucose tolerance was enhanced in each WT and Gpr120 KO mice fed PUFA HFD when compared with SAT HFD (Fig. 5B). In WT mice, blood glucose area under the curve (AUC) was 1714.110.5 on PUFA HFD and 2151.403.five on SAT HFD (p,0.05), and in Gpr120 KO mice, blood glucose AUC was 1532.57.0 on PUFA HFD and 1817.10.6 on SAT HFD (p,0.01). The insulin response measured as AUC was significantly reduce following the glucose challenge in each genotypes when fed the PUFA HFD as when compared with the SAT HFD. In WT mice, blood insulin AUC was 257.63.4 on PUFA HFD and 683.507.6 on SAT HFD (p,0.01), and in Gpr120 KO mice, blood insulin AUC was 304.60.6 on PUFA HFD and 554.04.7 on SAT HFD (p,0.05). The 15 minute insulin response in Gpr120 KO mice on PUFA HFD was additional STING Inhibitor site marked and correlated using a trend towards reduce blood glucose levels at 30 minutes in the Gpr120 KO mice when compared with WT mice on PUFA HFD (Fig. 5B).Tissue weights and histologyFinal body weight was 18 reduce in WT mice and 12 reduced in Gpr120 KO mice on PUFA HFD as in comparison to the corresponding groups on SAT HFD (Table two). Interestingly, the relative weights of epididymal and retroperitoneal fat depots tended to be higher in WT animals and was significantly higher in Gpr120 KO animals on PUFA HFD as in comparison with these on SAT HFD. Having said that, there was no impact on diet regime or genotype on relative brown adipose tissue (BAT) weights. The relative liver weight was about 40 lower in both WT and Gpr120 KO animals on PUFA HFD. Epididymal white adipose tissue (WAT) was analysed in terms of macrophage content. No considerable variations in Mac2 quantified staining have been observed in between PUFA HFD and SAT HFD fed mice. In WT mice, Mac2 area was 1.14.23 on PUFA HFD and 0.98.34 on SAT HFD, and in Gpr120 KO mice, the Mac2 area was 0.98.21 on PUFA HFD and 0.80.22 on SAT HFD (representative slides shown in S3 Fig.). WAT tissue was also double stained with Perilipin and Mac2 to understand how the different pattern of immune markers correlated with dead adipocytes (Fig. 6). As expected, adipose tissue from mice fed SAT HFD displayed high quantity of `crown like’ structures (CLS) surrounding perilipin-free lipid droplets (Fig. 6 and S3 Fig.). Interestingly, staining with the WAT macrophages in mice fed the PUFA HFD revealed the presence of equivalent numbers of immunopositive macrophages but these displayed a various pattern of Mac2-staining as multinuclear giant cells aggregation.