4.3. Cell Viability Assay

The effects of flaccidoxide-13-acetate on the T24 and RT4 cell lines were evaluated by MTT assay. Cells (1 × 10⁵ cells/well) were seeded in 96 well plates. The cells were treated with various concentrations of flaccidoxide-13-acetate (5, 10, 15, 20, and 25µM). After 24 h of incubation, MTT solution (0.5 mg/mL in PBS) was added to each well. The plates were incubated for 4 h at 37 °C, after which the culture medium was removed and the cells were dissolved in 200 μL DMSO. The absorbance was measured at 595 nm using a microplate ELISA reader (Bio-Rad, Hercules, CA, USA) and DMSO was used as the control. Samples were analyzed and all experiments were repeated three times.