Triglycerides e test

Blood samples (0.8 mL) were collected from the retro-orbital sinus of fasting mice and the level of glucose was measured by the glucose oxidase method (Model 1500; Sidekick Glucose Analyzer; YSI Incorporated, Yellow Springs, OH, USA). Plasma triglycerides (TG), total cholesterol (TC), and free fatty acids (FFA) were analyzed using commercial assay kits according to the manufacturer's directions (Triglycerides-E test, Cholesterol-E test, and FFA-C test, Wako Pure Chemical, Osaka, Japan).

Fasting blood glucose levels were determined by collecting blood samples (> 20 μL) drawn from retro-orbital sinuses. Approximately 20 μL of fresh blood sample was quickly placed on tinfoil, and the blood glucose level was read using a machine (Model 1500; Sidekick Glucose Analyzer; YSI Incorporated, Yellow Springs, OH, USA) by the glucose oxidase method. Blood TG and TC levels were determined using commercial assay kits in accordance with the manufacturer’s directions (Triglycerides-E test and Cholesterol-E test, Wako Pure Chemical, Osaka, Japan). Aliquots of plasma samples (> 25 μL) were collected for insulin, adiponectin, and leptin assays using enzyme-linked immunosorbent assay (ELISA) kits (mouse insulin ELISA kit, Mercodia, Uppsala, Sweden; adiponectin ELISA kit, Crystal chem; mouse leptin ELISA kit, Morinaga, Yokohama, Japan) as described in previous studies [57 (link),58 (link)]. The percentage of HbA1c was determined using a Hemoglobin A_1C kit (BioSystems S.A., Barcelona, Spain).

Blood samples (0.8 mL) were collected from the retro-orbital sinus of fasting mice and the level of glucose was measured by the glucose oxidase method (Model 1500; Sidekick Glucose Analyzer; YSI Incorporated, Yellow Springs, OH, USA). Plasma triglycerides (TG), total cholesterol (TC), and free fatty acids (FFA) were analyzed using commercial assay kits according to the manufacturer's directions (triglycerides-E test, cholesterol-E test, and FFA-C test, Wako Pure Chemical, Osaka, Japan).

The blood samples (about ≈150–200 μL) were acquired from the retro-orbital sinus of mice after 10 h fasting (8:00 a.m.), and the levels of blood glucose were measured by automatic analyzer (model 1500; Sidekick Glucose Analyzer; YSI Inc., Yellow Springs, OH, USA), which is based on glucose oxidase method. The concentrations of plasma TG, TC, and FFA were measured by commercial kits (Triglycerides-E test, Cholesterol-E test, and FFA-C test, Wako Pure Chemical, Osaka, Japan). The levels of insulin, leptin, and adiponectin were measured by enzyme-linked immunosorbent assay (ELISA) by commercial kits (insulin ELISA kit, Sibayagi, Gunma, Japan and leptin ELISA kit, Morinaga, Yokohama, Japan; adiponectin ELISA kit, Crystal Chem, Inc., Suite A Downers Grove, IL, USA).

Blood samples were collected from the retro-orbital sinuses of 12 h fasted mice. Blood glucose levels were determined by the glucose oxidase method (Model 1500; Sidekick Glucose Analyzer; YSI Incorporated, Yellow Springs, OH, USA). The levels of TG, TC, and free fatty acids were determined using commercial assay kits in accordance with manufacturer’s directions (Triglycerides-E test, Cholesterol-E test and FFA-C test, Wako Pure Chemical, Osaka, Japan). Insulin and leptin levels were measured by enzyme-linked immunosorbent assay (ELISA) (mouse insulin ELISA kit, Mercodia, Uppsala, Sweden; mouse leptin ELISA kit, Morinaga, Yokohama, Japan).