Cell Viability Assay with Cyquant Reagent

Cell survival assay using Cyquant reagent (Invitrogen, C7026) was performed as described (19 (link)). Briefly, 1–4 x 10⁵ cells/mL were seeded in 96 well plates overnight, before treatment with either vehicle or drug for 48–72 h. Cells were harvested, washed and frozen at -80°C overnight. After thawing, Cyquant reagent was added and fluorescence intensity was measured in a spectrometer at 485 nm. Relative percent survival was calculated by dividing the reading from drug-treated cells by the reading from vehicle-treated cells. Median-dose effect analysis and ED50 calculation (50% reduction in cell survival) was performed using CompuSyn software (ComboSyn, Inc.), as described previously (21 (link)).