Caspase-3 activity assay

Caspase-3 activity was measured by using a CaspGLOW Red Active caspase-3 activity kit from Bio Vision (K-193) as per the manufacturer's instructions. Briefly, 1 × 10⁶ cells were stained with 1 µL of Red-DEVD-FMK for 30 minutes at 37°C and 5% CO₂. Cells were washed thrice using wash buffer and analysis was done using BD FACS-CantoII (Becton & Dickinson) equipped with a 488-nm Coherent Sapphire Solid State laser. Red (564–606 nm) fluorescence emission from 10⁴ cells was measured after illumination with blue (488 nm) excitation light. Data were analyzed using Summit software V5.2.1.12465.