Blood samples were collected from retro-orbital sinus of conscious mice, and used to determine blood glucose levels (OneTouch Basic glucometer or AlphaTRAK monitor as indicated). Plasma was collected in EDTA tubes. Body composition was determined using the EchoMRI apparatus. Glucose tolerance tests (GTT) were conducted in 12-hour fasted mice following IP injection of 2 g/kg glucose. Plasma cholesterol, triglycerides, and NEFA levels were measured using the Olympus AU400e Chemistry Analyzer (Olympus America, Inc; Center Valley, PA). Insulin and adiponectin values were determined by ELISA (Crystalchem and Millipore respectively). Plasma FGF21 levels were measured using an in-house ELISA [15]. Recombinant murine/human FGF21 and murine leptin were generated as previously described [15], [19].
