Wako l type tcho

Liver tissues were collected by surgical manipulation, and fixed using 10% buffered formalin solution (Histo-Fresh, Pharma, Tokyo, Japan). Total lipids were extracted using a mixture of methyl tert-butyl ether (Wako Pure Chemical, Osaka, Japan) and methanol (Wako Pure Chemical, Osaka, Japan), as previously reported [37 (link)]. The dried lipid residues were dissolved in 20 µL of isopropanol containing 10% Triton X-100 (Nakarai Tesque, Kyoto, Japan). TG and TCHO were measured using Wako L-type TG and Wako L-type TCHO (Wako Pure Chemicals, Tokyo, Japan) assay kits according to the manufacturer’s protocol.

NSY/HOS mice, a type 2 diabetes mellitus strain^{42 (link)}, was purchased from Hoshino Laboratory Animals (Saitama, Japan) and housed on a 12-h light/dark cycle at the Institute of Laboratory Animals at Mie University. Six-month-old male mice were assigned to three groups with six mice, housed individually, and were fed either the CE-7 normal diet (ND; CLEA Japan, Tokyo, Japan), high-fat diet (Test Diet 58Y1; TestDiet, Richmond, IN, USA) or a high-fat diet (HFD) supplemented with RS (250 mg/Kg BW) for 4 weeks to induce obesity. The compositions of ND and HFD are described in Table S2. During the feeding experiment, body weight, food intake, and faecal weight were measured once per week. Mice were subjected to fasting for 14 h before blood sampling to assess the fasting blood glucose levels. Blood glucose was measured using a hand-held glucometer (Glutest Neo Super; Sanwa Kagaku, Nagoya, Japan). The plasma levels of triacylglycerol (TG), low-density lipoprotein cholesterol (LDL-C), and total cholesterol (TCHO) were measured using Wako L-type TG, Wako L-type LDL-C, and Wako L-type TCHO (Wako Pure Chemicals, Osaka, Japan) assay kits according to the manufacturer’s protocol. The mice were euthanized with CO₂ gas, and then the organ samples were collected and subsequently dissected for analysis.

At the end of the feeding experiment, zebrafish were deprived of food overnight to provide fasting levels of blood glucose, and blood was withdrawn from the dorsal artery by a heparinized glass capillary needle (GD-1, Narishige, Tokyo, Japan) as previously reported [35 (link),36 (link)]. Blood glucose was measured using a hand-held glucometer (Glutest Neo Super, Sanwa Kagaku, Nagoya, Japan). The plasma levels of triacylglycerol (TG), low-density lipoprotein cholesterol (LDL-C) and total cholesterol (TCHO) were measured using Wako L-type TG, Wako L-type LDL-C, and Wako L-type TCHO (Wako Pure Chemicals, Osaka, Japan) assay kits according to the manufacturer’s protocol.