Cell Viability Assay for Drug Screening

Cells were seeded in 96-well plates and treated in triplicate for 72 h with 0–10 μM concentrations of the relevant drug diluted in the appropriate growth medium (please see above). Cell viability was measured using CellTiter-Glo® reagent per the manufacturer’s instructions (Promega, Madison, WI, USA) and ATP measurements were read using an Envision reader (PerkinElmer, San Jose, CA, USA). Cell viability measurements were corrected to the day 0 viability (day of treatment) as measured by CellTiter-Glo® and plotted relative to the dimethyl sulfoxide control (100% viability).^{48 (link)} With this method, cell growth after treatment can be compared to that before treatment (i.e., zero percent growth represents cell stasis and negative percent growth represents cell death). Three replicate wells per treatment were used, such that, at 5% standard deviation, viability differences of >9% should be detectable with a power of 80%. Data shown represent three independent experiments.