Thermo labsystem multiscan spectrum

For the detection of cytotoxicity of asiatic acid, BMMs were cultured at 6×10³ cells per well in complete α-MEM with M-CSF for 12 h incubation (condition as 5% CO2 at 37°C). Different concentrations of asiatic acid were then added to BMMs with the complete α-MEM with M-CSF for total two days. After that, 20 µl MTS solution (Promega, Sydney, Australia) was added to each well for 2 h. The optical density (OD) manifested by the MTS solution was measured by a BMG plate reader (Thermo Labsystem Multiscan Spectrum, Thermo Fisher, Waltham, MA, USA) with 490 nm wavelength.

The potential cytotoxicity of Bajijiasu was assessed using an in vitro cell death detection assay. BMMs were plated at a concentration of 6 × 10³ cells per well in 96-well plates in 5% CO₂ at 37 °C. After an overnight incubation to allow adherence, BMMs were treated with complete α-MEM medium with M-CSF (25 ng/mL) and varying concentrations of Bajijiasu (0.05, 0.1, 0.2, 0.4, 0.8 mM). After culturing for 48 h 20 μL MTS ((3-4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) solution was added to each well and further incubated for 2 h in 5% CO₂ at 37 °C. The optical density (OD) was measured at 490 nm wavelength using a BMG plate reader (Thermo Labsystem Multiscan Spectrum, ThermoFisher, Waltham, MA, USA).