Apoptosis Induction in PC3 Cells

PC3 cells (2×10⁵ per well) were seeded into a 6-well plate in RPMI 1640 medium supplement with 10% FBS, 1% pen/strep and incubated overnight in 5% CO₂ incubator at 37°C and 95% humidity. Then, the cells were treated with 5-FU (0.75 μM) and rutin (700 μM) or combination of 5-FU and rutin (0.75 μM and 700 μM respectively) for 48 hours. Subsequently, the cells were collected by trypsinization and washed with PBS and stained by Annexin V/propidium (BD Biosciences) based on the manufacturer’s instructions for 25 minutes at room temperature in a dark place.^{25 (link)} The stained PC3 cells were analyzed using a flow cytometer (FACScan; Becton Dickinson Immunocytometry Systems, San Jose, CA, USA). Experiments were performed in triplicates.