2.3. MTT Cell viability assay

96-well plate was seeded with 3×10³ PC3 cells in 100 μL of complete medium (RPMI-1640 with 10% FBS and antibiotics) per well. The plate was incubated at 37°C with 5% CO₂ until 50% confluent. The old media was removed and the cells were treated with drug media containing varying concentrations of drugs (NCX4040, Aspirin and DETA NONOate), with or without inhibitors (NAC, GSH, Catalase, DETDC and PTIO) and untreated cells served as controls. The plates were incubated at 37°C with 5% CO₂ for the duration of the treatment period (48hr). Afterwards, MTT dye was added to each well and incubated for 2–3 hrs. After incubation, solubilization solution was added to dissolve the crystals. The plates were read at 570nm under a spectrometer (Biotech Synergy 2). The percentage of cell viability calculated as follows.