Cell Proliferation Assay for Maduramicin

Cell proliferation was also evaluated using One Solution Cell Proliferation Assay Kit (Promega), as described [24] (link). Briefly, cells suspended in the growth medium were seeded in a 96-well plate at a density of 1×10³ cells/well (in triplicates) and grown overnight at 37°C in a humidified incubator with 5% CO₂. The next day, maduramicin (0–1 µg/ml) was added. After incubation for 24–72 h, each well was added 20 µl of one solution reagent and incubated for 1 h. Cell proliferation was determined by measuring the OD at 490 nm using a Wallac 1420 Multilabel Counter (PerkinElmer Life Sciences, Wellesley, MA, USA). Cells treated with the vehicle (DMSO) alone were used as a control.