Accu check test strips

Glucose and insulin tolerance tests were performed on overnight fasted, or 4-h fasted mice, respectively. Blood glucose levels were measured over a period of 3 h after injection of glucose (Roth X997.2) 2 mg/g body weight or insulin 0.5 µ i.U./g body weight (Huminsulin Normal 100 Lilly, HI0210) using an Aviva Accu-Check, and Accu-Check test strips (Roche). Fasted serum insulin levels were determined using Ultrasensitive mouse insulin ELISA (Mercodia 10-1249-01). Serum non-esterified fatty acids (NEFA) were measured on serum from fed mice (NEFA-HR(2) Assay, Wako Chemicals Europe). ALT (Sigma, MAK052) and AST (Sigma MAK055) measurements were made on serum isolated from fed mice. Lipase activity assay was performed on frozen eWAT isolated from obese mice according to the manufacturer’s instructions (Sigma, MAK046). Corticosterone was measured using mass spectrometry as described before^{81 (link)}.

The Intraperitoneal Glucose Intolerance Test (ipGTT) was performed after 5–6 h of food deprivation by administration of an intraperitoneal injection of D-glucose (2 g/Kg body weight). Glucose in tail vein blood was measured at the indicated time points after injection using a clinical glucometer and Accu-Check test strips (Roche Diagnostics, Switzerland). Plasma samples were obtained by blood centrifugation and kept at −80 °C for subsequent insulin determination by ELISA.

In order to assess whole-body glucose tolerance, intraperitoneal glucose tolerance tests (IPGTT) were performed after 6 h food deprivation in Atoh8 Δ^panc and wild-type mice. The IPGTT was performed by administration of an injection of D-glucose (2 g/Kg body weight), and blood samples were collected from tail vein at 0, 15, 30, 60 and 120 minutes after injection. Glycaemia was measured at the same time points using a clinical glucometer and Accu-Check test strips (Roche Diagnostics, Switzerland). Plasma was obtained by blood centrifugation and kept at -80°C for insulin determination using a mouse insulin ELISA kit (Mercodia, Uppsala, Sweden).