All experiments were approved by the McMaster University Animals Ethics Committee and conducted under appropriate Canadian guidelines for animal research. Mice lacking adipocyte AMPK β1β2 (iβ1β2AKO) along with littermate controls were generated as previously described [23] (link). Tamoxifen (Cayman Chemical; MI, USA), dissolved in sunflower oil, was given via oral gavage for five consecutive days in adult males to induce the deletion of adipocyte AMPK β1β2 after 10 weeks of a high fat diet (HFD, 45% kcal fat) (D12451, Research Diets; New Brunswick, New Jersey). At week 14 of HFD, control and iβ1β2AKO mice were anesthetized with Avertin (2,2,2-tribromoethanol dissolved in methyl-2-butanol, Sigma-Aldrich; ON, Canada; 0.5 mg/g body) and implanted dorsally with two week mini-osmotic pumps (1002, Alzet; CA, USA) delivering 0.35 mg/kg/day of native FGF21 or 0.9% sodium chloride (saline) and incisions closed with perma-hand silk suture (Ethicon; ON, Canada). Food and water were provided ad libitum and mice were housed at 23 °C with a 12-hour light/dark cycle.
Male ACC1-S79A and ACC2-S212A knock-in mutation mice (ACC DKI) were as previously described and were put on a 45% HFD at 3–4 months of age [31] . At week 10 of HFD, wildtype and ACC DKI mice received subcutaneous injections of long-acting FGF21 (PF-05231023) at 10 mg/kg twice a week for 2 weeks.
Male ACC1-S79A and ACC2-S212A knock-in mutation mice (ACC DKI) were as previously described and were put on a 45% HFD at 3–4 months of age [31] . At week 10 of HFD, wildtype and ACC DKI mice received subcutaneous injections of long-acting FGF21 (PF-05231023) at 10 mg/kg twice a week for 2 weeks.