Plasma insulin (ALPCO), C-peptide (ALPCO), DPP4 (Thermo Fisher Scientific), active and total GLP-1 (ALPCO), GIP (Crystal Chem), and glucagon (Mercodia) levels were measured by enzyme-linked immunosorbent assay (ELISA) in accordance with the manufacturers’ instruction. To prevent degradation of active GLP-1 or active GIP by DPP4 during or after sample collection, DPP4 inhibitor (Millipore, catalog no. DPP4-010) was briefly added to each tube after sample collection. Plasma and tissue DPP4 activity was measured with a fluorometric assay kit (BioVision). Pancreatic insulin content was determined as described previously (60). Briefly, whole pancreas from each mouse was ground in 10 ml of acidic ethanol using a Polytron tissue homogenizer (Kinematica), and insulin levels were measured by ELISA in the tissue lysates. Total insulin content was calculated by multiplying the lysate insulin concentration, dilution factor, and total volume of the tissue lysate.

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