Prestoblue hs cell viability dye

NSCLC cells were seeded onto 24 well culture plates at a density of 1.3 x 10⁵ cells per well (~60% confluency) on Day 0. On Day 1, inhibitors were added to culture media and mixed thoroughly. On Day 3, culture media was aspirated, cells were washed with 1 mL of PBS, and subsequently incubated with fresh media and PrestoBlue HS cell viability dye (ThermoFisher P50200); PrestoBlue HS cell viability dye was added at a 1:10 (volume:volume) ratio according to manufacturer instructions. PrestoBlue HS dye was also incubated with cell-free, media only controls to account for background signal from culture media. After PrestoBlue HS addition, culture plates were incubated at 37°C for two hours. Following incubation, PrestoBlue HS fluorescence signal was quantified using a SpectraMax M2 microplate reader. Resulting signal was background corrected and is reported as a ratio of 560/590 nm fluorescence.