C3h10t1 2 cell line

The C3H10T1/2 cell line was obtained from the RIKEN cell bank (Tsukuba, Japan). Cells were maintained in Dulbecco’s modified Eagle’s medium (GIBCO, Tokyo, Japan), supplemented with 10% fetal bovine serum, in polystyrene dishes at 37°C under 5% CO₂. Differentiation medium was prepared by supplementing the maintenance medium with 0.1 μM dexamethasone (Sigma-Aldrich, St. Louis, MO), 0.17 mM ascorbic acid (Sigma-Aldrich), 10 μg/ml human transferrin (Roche Molecular Biochemicals, Mannheim, Germany), 3.0 × 10^-8 M sodium selenite (Sigma-Aldrich). Maintenance medium or differentiation medium was changed twice a week without damaging the gels.

C3H10T1/2 cell line was obtained from the RIKEN Cell Bank (Tsukuba, Japan). C3H10T1/2 cells were plated in 6-well plate in Dulbecco’s Modified Eagle Medium (DMEM, Cat. #11885-084, Gibco, Thermo Fisher Scientific, MA, United States) containing 10% fetal bovine serum (FBS, Cat. #7524, Nichirei Biosciences, Tokyo, Japan) and 100 U/mL penicillin, and maintained in a humidified incubator at 37°C under a 5% CO₂ atmosphere into a confluent. For the differentiation procedure, confluent C3H10T1/2 cells were induced to differentiate into adipocytes in a differentiation medium supplemented with 1 μM Rosiglitazone (Cat. #R0106, Tokyo Chemical Industry, Tokyo, Japan) for 48 h. Rosiglitazone was dissolved in dimethyl sulfoxide (DMSO, Cat. #D8418, Sigma-Aldrich, St. Louis, MO, United States). 1 mM Pro-Hyp (Cat. #4001630, Bachem, Bubendorf, Switzerland) was diluted in the DMEM medium and co-treated with Rosiglitazone for 48 h.