Cobas b 101 instrument

At the end of the treatment, blood was collected from the tail tip of each mouse and fasting blood glucose levels were determined using Glucocard™ blood glucose meter (Menarini, Italy). Then, animals were anaesthetized with an intraperitoneal injection of aqueous urethane solution 30% w/w. Intracardiac blood was collected in tubes with the anticoagulant EDTA (BD Vacutainer), and complete lipid panel (triglycerides, total cholesterol, HDL, and LDL) and glycated haemoglobin levels were measured with Cobas b 101 instrument (Roche Diagnostics, Switzerland).
Data analysis. Blood samples were collected from 15 mice per group and results were expressed as mean ± SEM. Cardiovascular risk was expressed as ratio between total cholesterol and HDL cholesterol levels. One-way ANOVA followed by Bonferroni's post hoc test was selected as statistical analysis, and the difference between groups was considered statistically different when p < 0.05.

Plasma glucose concentration was measured using a hexokinase method at a commercial laboratory (Melbourne Pathology, Collingwood, Victoria, Australia) with a CV < 1.9%. Fasting triglycerides were analysed using a Cobas b 101 instrument (Roche Diagnostics Ltd., Basel, Switzerland). Plasma insulin, connecting peptide (C-peptide) and tGLP-1 concentrations were measured using a Luminex Analyzer (MAGPIX^®; Human Metabolic Hormone Magnetic Bead Panel, EMD Millipore, Burlington, MA, USA) following manufacturer’s instructions with intra-assay CV’s of 9.8%, 6.8% and 11.4%, respectively.