Fuji dri chem 4000i analyzer

We determined creatine kinase (CK) using Fuji Dri-Chem 4000i-Analyzer (FujiFilm, Tokyo, Japan) and Fuji Dri-Chem Slides (CPK-P III, FujiFilm) according to the protocol provided by the company.

Lipids from liver and feces were extracted using a modified Folch method [18] . The isolated lipid fraction was used to measure TOTAL-CHOL and TG concentrations using commercially available kits (Asan Pharm, Seoul, Korea). Serum TOTAL-CHOL, HDL-CHOL, and TG levels were also measured with commercially available kits (Asan Pharm, Seoul, Korea). All determinants were obtained by absorbance at 550 nm using a microplate reader (Molecular Device, Sunnyvale, CA, USA). LDL-CHOL was calculated using the Friedewald formula [19] (link) (LDL-CHOL (mg/dl) = TOTAL-CHOL – HDL-CHOL – TG/5). Atherogenic indices were obtained by the Haglund method (Atherogenic index = (TOTAL-CHOL – HDL-CHOL)/HDL-CHOL) [20] . Bile acid was extracted using the DeWael method [21] (link). Extracted fecal bile acid was measured using a kit from Bioquant (Nashville, TN, USA). Serum glucose levels were measured using the Fuji DRI-CHEM 4000i analyzer (Fuji Film Co., Tokyo, Japan).

Glucose, insulin and pyruvate tolerance tests were performed after an overnight fast (∼15 h) or morning fast (3 h) with free access to water. Mice were weighed before an intra-peritoneal injection of glucose (2 g/kg, Sigma-Aldrich), insulin (0.75 U / kg, Novo Nordisk, Denmark) or pyruvate (2 g/kg, Sigma-Aldrich), respectively. Blood samples from live animals were collected from the tail tip and blood glucose level was measured with a glucose meter (Accu-Chek, Roche, Switzerland) at regular intervals as indicated in the figure legends.
Levels of total cholesterol (free cholesterol + cholesterol esters) and triglycerides present in serum were measured using the Fuji Dri-Chem 4000i analyzer (FUJIFILM Corporation, Tokyo, Japan). Blood was collected from the harvested animals via heart puncture.