Mak052

Plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured using colorimetric enzymatic assay kits (cat# MAK052 (ALT) and cat# MAK055 (AST), Millipore Sigma, St. Louis, MO, USA), according to the manufacturer’s instructions.

Plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured using colorimetric enzymatic assay kits (cat# MAK052 (ALT) and cat# MAK055 (AST), Millipore Sigma, St. Louis, MO), according to the manufacturer’s instructions.

Glucose and insulin tolerance tests were performed on overnight fasted, or 4-h fasted mice, respectively. Blood glucose levels were measured over a period of 3 h after injection of glucose (Roth X997.2) 2 mg/g body weight or insulin 0.5 µ i.U./g body weight (Huminsulin Normal 100 Lilly, HI0210) using an Aviva Accu-Check, and Accu-Check test strips (Roche). Fasted serum insulin levels were determined using Ultrasensitive mouse insulin ELISA (Mercodia 10-1249-01). Serum non-esterified fatty acids (NEFA) were measured on serum from fed mice (NEFA-HR(2) Assay, Wako Chemicals Europe). ALT (Sigma, MAK052) and AST (Sigma MAK055) measurements were made on serum isolated from fed mice. Lipase activity assay was performed on frozen eWAT isolated from obese mice according to the manufacturer’s instructions (Sigma, MAK046). Corticosterone was measured using mass spectrometry as described before^{81 (link)}.

Selenium content was measured in serum by atomic absorption spectrophotometer (AA6501, Shimadzu Ltd., Japan). Plasma samples were used for measuring of aspartate amino transferase (AST), alanine glutamyl transferase (ALT), and creatinine calorimetrically by diagnostic kits (MAK055, MAK052, and C4255, respectively) manufactured by Sigma-Aldrich.

Trunk blood was collected right after the animal sacrifice and allowed to clot for 30 min at room temperature, followed by 5000 g centrifugation for 10 minutes at 4° C to collect the serum fraction [19 (link), 73 (link)]. Serum creatinine and ALT levels were measured using the following assay kits respectively: DICT-500 (BioAssay Systems) and MAK052 (Sigma-Aldrich). Assays were performed according to manufacturer instructions.

Red blood cell (RBC) counts were established as specified by Brown (1976). Hemoglobin (Hb) concentration was quantified according to [17 ]. The serum concentrations of creatinine, urea alanine transaminase (ALT) and aspartate transaminase (AST) were calculated using standard kits (Sigma-Aldrich; MAK052, MAK055, MAK080 and MAK006, respectively). C-reactive protein (CRP) levels were evaluated according to the guidelines of a commercial kit (AG723-M; Sigma-Aldrich).