Luminescent cell viability assay

Concentration-response curves and IC₅₀ values were calculated using the Cell Titre-Glo^® Luminescent Cell Viability Assay kit (Promega, USA). Briefly, cells were plated in 96-well plates for 24 hours followed by Cud C treatment for 72 hours Luminescence was measured using SpectraMax M3 Multi-Mode Microplate Reader (Radnor, USA). The experiments were validated using CRC treated with 5-fluorouracil (Sigma-Aldrich, USA), the clinically used anti-cancer agent. Cell viability was expressed as a percentage of the vector-treated control.