Advantage 2

Mouse vesiculin was synthesized in-house by the peptide synthesis group as per their published procedure in which the A- and B-peptide chains were first synthesized separately with protected cysteine residues before using regioselective disulfide bond formation;^{21 (link)} stocks were prepared at a final concentration of 1 μg/μl in 16 mg/ml glycerol; 0.1% DDM. Insulin (Actrapid® Recombinant human-insulin 100 IU/ml; 3.47 μg/μl) was from Novo Nordisk (Bagsvaerd, Denmark). Both were diluted in physiological saline to the desired final concentrations. Blood glucose was measured in tail-vein blood (Advantage II; Roche). Core body temperature was recorded using a rectal probe (RET-3) coupled to a thermometer (Physitemp Instruments Inc).

Mice were fasted for 8 hours before the glucose tolerance test. A small fragment of tail tissue was cut for blood collection and glucose was quantified using an Advantage II glucometer (Roche). Glucose measurements were carried out before and at 15, 30, 60, and 120 minutes after intraperitoneal injection of glucose 1 g/kg body weight.