Mice were bred and maintained in the Laboratory Animal Research facility at the University of Kansas Medical Center in rooms under a 12-hr light/dark cycle. All protocols were approved by the IACUC. All experiments used 10–16 week old male mice and all mice were sacrificed within a 30-min period in the morning. In addition, all treatments were repeated twice.
Activation of Fxr in vivo was achieved by treatment with an Fxr synthetic agonist, GW4064, at 150 mg/kg. GW4064 or vehicle was administered by oral gavage at 6 PM, followed by a second administration at 8 AM next morning. Two hrs later, liver and ileum were harvested. The generation of purified Fgf15 has been reported previously.15 (link) Fgf15 protein was injected to mice through tail vein at a dosage of 10 μg/kg. Two hrs or indicated time points (for time course study) after the injection, livers were collected.
The total bile-acid pool size was determined by measuring bile acids of the small intestine, gallbladder, liver, and their contents. Ten-sixteen week-old mice were fed a chow diet with 2% cholestyramine for 10 days. Total bile acids were determined using a kit from Bio-quant Kits Inc. (San Diego, CA), and the pool size was expressed as micromoles of bile acids/100 g of body weight.
Activation of Fxr in vivo was achieved by treatment with an Fxr synthetic agonist, GW4064, at 150 mg/kg. GW4064 or vehicle was administered by oral gavage at 6 PM, followed by a second administration at 8 AM next morning. Two hrs later, liver and ileum were harvested. The generation of purified Fgf15 has been reported previously.15 (link) Fgf15 protein was injected to mice through tail vein at a dosage of 10 μg/kg. Two hrs or indicated time points (for time course study) after the injection, livers were collected.
The total bile-acid pool size was determined by measuring bile acids of the small intestine, gallbladder, liver, and their contents. Ten-sixteen week-old mice were fed a chow diet with 2% cholestyramine for 10 days. Total bile acids were determined using a kit from Bio-quant Kits Inc. (San Diego, CA), and the pool size was expressed as micromoles of bile acids/100 g of body weight.
