Dose-response and Invasion Assay Protocol

Dose–response curves and cell viability were assessed using alamarBlue® cell viability assay (Serotec, Oxford, UK) 5 and 10 days post-treatment, respectively^{37 –39 (link),53 (link)}. Dose–response curves for each cell line were generated from a mean of triplicate experiments using GraphPad Prism (GraphPad Software, Inc; USA) and IC₅₀ values were computed from 10-point titration curves ranging from 10⁻⁴ to 10² μM. For invasion assay, 50,000 cells were added to the upper compartment of the Corning® BioCoat™ Matrigel® invasion chamber (BD Biosciences, San Jose, CA) and 2% fetal calf serum was supplemented into the lower compartment. Cells were incubated for 24 h and the lower surface was subsequently stained with 0.005% crystal violet (Sigma-Aldrich). The number of cells from five random fields having migrated to the bottom chamber was counted.