*2.4. Cell Proliferation and Cytotoxicity Assay*

The cytotoxicity of Rg1 was analyzed by using the WST-1 Cell Proliferation and Cytotoxicity Assay Kit (Beyotime). Marc-145 cells (2 <sup>×</sup> 104 cells per well) and PAMs (1 <sup>×</sup> 105 cells per well) were seeded in 96-well plates and grown at 37 ◦C. Cells were incubated with medium supplemented with different concentrations of Rg1 and further incubated for 48 h. The absorbance of each well was read at 450 nm with a reference wavelength of 630 nm using a microplate reader (Thermo Fisher Scientific, MA, USA).

For the cell proliferation assay, Marc-145 cells (5 <sup>×</sup> <sup>10</sup><sup>3</sup> cells per well) were seeded in 96-well plates and cultured without FBS at 37 ◦C for 12 h and then treated with indicated concentrations of Rg1. After 24 h of Rg1 incubation, the relative proliferation was evaluated using the WST-1 kit according to the instructions. The absorbance was read at 450 nm with a reference wavelength of 630 nm using a microplate reader (Thermo Fisher Scientific, MA, USA).
