Cell proliferation activity assay

The effect of Ma on proliferation in Caco-2 colon-cancer cells was measured using the MTT assay, which is based on the ability of live cells to cleave the tetrazolium ring, thus producing formazan, which absorbs at 570 nm. Caco-2 cells (15 x 10³) were grown in a 96-well plate and incubated with MA (0–100 μg/mL). Following 72 h, 100 μL of MTT solution (0.5 mg/mL) was added to each well. Following 2 h incubation, the cells were washed twice with PBS and the formazan was resuspended in 200 μL DMSO. Relative cell viability was measured by absorbance at 550 nm on an ELISA plate reader (Tecan Sunrise MR20-301, TECAN, Austria).