Tg e test

Plasma levels of triglyceride (TG), aspartate aminotransferase (AST), and alanine transaminase (ALT) were measured using the TG E-test (Wako Chemical, Osaka, Japan) and the transaminase CII-test (Wako Chemical), respectively. High-density lipoprotein cholesterol (HDL-chol) and low-density (LDL-chol) and very low-density lipoprotein-cholesterol (VLDL-chol) were assayed using HDL-C and LDL-C/VLDL-C Quantification Kit (BioVision, Milpitas, CA, USA).

Whole venous blood was obtained from the tail veins of the mice, and their glucose levels were measured using Medisafe Fit Smile (Terumo, Tokyo, Japan). Liver triglycerides (TGs) were extracted by homogenizing 50 mg of liver tissue in 1.25 mL of chloroform/methanol (chloroform:methanol = 2:1 (v/v)). After adding 0.4 mL of chloroform and 0.4 mL of water, the samples were vortexed and centrifuged for 3 min at 3,000 rpm. The lower layer was collected, evaporated, and dissolved with 10% Triton X-100 in isopropanol. TG concentrations were measured using the TG E-test (Wako).

Blood samples were collected from the tail vein and immediately used to determine glucose level using a glucometer (NovoAssist Plus, Novo Nordisk, Tokyo, Japan). Serum insulin level was measured using enzyme-linked immunosorbent assay kits (Lebis-insulin-mouse, Sibayagi, Gunma, Japan). Serum triglyceride (TG) and total cholesterol (TC) levels were measured using TG E-test and T-Cho E-test kits (Wako Pure Chemicals, Osaka, Japan), respectively. Total liver lipids were extracted using the previously described protocol with minor modifications [24 (link)]. Briefly, liver samples (100 mg) were homogenized in 1 mL of hexane/isopropyl alcohol (3:2, v/v). The homogenized samples were centrifuged, and the supernatant was evaporated under reduced pressure. The dried samples were then resuspended in 10% triton X-100 prepared in isopropyl alcohol, and the lipid levels were assayed using commercial kits. FGF21 levels in serum and culture medium were measured using mouse FGF-21 DuoSet ELISA (R&D SYSTEMS, Minneapolis, MN, USA).

Liver and serum triglyceride (TG), total cholesterol (TC) levels, and serum aspartate aminotransferase (AST) and alanine transaminase (ALT) activities were measured using the TG E‐test, cholesterol E‐test, and transaminase CII‐test (all from Wako Pure Chemical Industries), respectively. The livers (100 mg) were homogenized (4,000 rpm for 3 min at 4°C) with a bead cell disruptor Micro Smash MS‐100R (TOMY SEIKO Co., Ltd.); lipids were extracted with chloroform‐methanol (2:1) and subjected to lipid analysis. The extract was dried under nitrogen gas and re‐dissolved in 2‐propanol containing 5% (v/v) Tween‐20 for TG and TC determination.