Cell Viability Assay

The viability of 3T3-L1 cells was assessed with the MTT assay as previously described [22]. Cells were grown in 96-well plates at a density of 2 × 10⁴ cells/well. After adhering overnight, the cells were stimulated with AS in various concentrations (0.005, 0.05, 0.5 or 5 in DMSO). The control group was treated with DMEM with 10% FBS and 0.1% DMSO. After incubation for 24h at 37°C in 5% CO₂, cells were washed three times with phosphate-buffered saline (PBS). MTT reagent (10 μl of a 1 mg/ml solution) and medium (100 μl) were added to each well, followed by incubation in the dark at 37°C for 4h. Formazan crystals were solubilized with DMSO, and the absorbance at 570 nm was measured on an Elx-800 microplate reader (BioTek, Winooski, VT, USA).