Celltiter glo luminescent assay kit

HEK293 cell viability was determined based on quantification of ATP using Cell Titer-Glo^® luminescent Assay kit (Promega, Madison, WI, USA), which signals the presence of metabolically-active cells. Briefly, HEK293 cells were seeded into 96-well plates at a cell density of 3.5 × 10⁴ cells per well, the plates were kept at 37 °C with 5% CO₂ overnight. On the following day, cardamonin was added to the cells from a top concentration of 90 μΜ. The cells treated in the absence of the test compound were the negative control. After incubated for 24 h, Cell Titer-Glo reagent was added to the cells and Luminescence was acquired on EnVision™ 2100 Multilabel Reader (PerkinElmer, Santa Clara, CA, USA).

HeLa, MCF-7, and A549 cancer cells were seeded at a density of 5000 cells/well and maintained in an incubator overnight at 37 °C with 5% CO₂. The compounds were suspended in a medium at final concentrations of 100 mg/mL in DMSO and analyzed in a decreasing dose curve from 50 to 100 μM. As a control, cells were treated with 1–2% DMSO. The number of viable cells in the culture was determined via the quantification of ATP, using the Cell Titer-Glo luminescent assay kit (Promega, Madison, WI, USA). Following the manufacturer’s instructions, the cells were plated in 96-well plates and treated 24 h later with the compounds for 48 h and concentrations, followed by the addition of a Cell Titer-Glo reagent. Luminescence was detected using a multi-well Synergy Mx scanning spectrophotometer (Bio-Tek, Winooski, VT, USA).
GIST cell lines were seeded in 96-well plates (10,000 cells/well) and treated with the compounds for the indicated concentrations. Cell viability was measured using the colorimetric WST-1 assay (Roche™ Diagnostics, Mannheim, Germany) upon 72 h of treatment according to the manufacturer’s protocol. Data were expressed as the mean ± standard deviation (mean ± SD) from three independent experiments.