4.3. Cell Viability Assay

Drugs were prepared with DMSO as a 100 mM stock solution. An amount of 200 µL of cells (1 × 10⁴ cells/mL) were seeded in 96-well plates. Cells were treated in the range of 0.01 µM, 0.1 µM, 1 µM, 10 µM, 100 µM, and 1000 µM of drugs. For blank wells, the solvent (DMSO) was used according to the maximum number of dilutions to exclude any effect of the solvent. All of the treatments were performed in triplicate and incubated for 96 h at 37 °C. The media was discarded and the cells were incubated with MTT solution at a final concentration of 0.5 mg/mL. After 4 h, the MTT solution was removed and formazan crystals were resuspended with DMSO. The absorbance was measured using a Multi scan Plus spectrometer at 550 nm (Lab-systems Group, UK), the IC₅₀ results were calculated with The Quest Graph™ IC₅₀ Calculator [44], and graphs were plotted by Excel (version 2406).