Mouse glucose assay

Plasma leptin and glucose levels were determined using Mouse Leptin ELISA kit (Crystal Chem, USA) and Mouse Glucose Assay (Crystal Chem, USA), respectively. Mouse Insulin ELISA Kits, Ultra‐Sensitive Assay (Crystal Chem, USA) was used to determine the plasma insulin level.
The weights of liver and previously dried caecum content samples were recorded, and the samples were homogenised in 1,5 ml of %NP40/ddH2O solution. After two repeated steps of heating (80°C–100°C for 3–5 min) and cool down (15 min at RT), all the samples were centrifuged at maximum speed. The supernatant, containing the lipids, was collected and diluted 1:10. Triacylglycerol (TAG) level in the plasma, in the liver extract and in the caecum content extract it was measured using Triglyceride Quantification Assay Kit (abcam, UK).
Leptin, glucose, insulin, and TAG levels detection were carried out in all the groups, in both experiments.

Plasma glucose was measured by Mouse Glucose Assay (Crystal Chem, Downers Grove, IL, USA). Serum insulin was measured using an Ultra Sensitive Mouse Insulin ELISA kit (Crystal Chem). Serum triglycerides (TG), non-esterified fatty acids (NEFA), and cholesterol were measured using respective tests from Wako Diagnostics (FUJIFILM Wako Diagnostics, Mountain View, CA, USA). Serum adiponectin and leptin were measured using the Mouse Quantikine ELISA kit for the respective analyte (R&D Systems, Minneapolis, MN, USA). All samples were analyzed according to the manufacturers’ instructions.