MTT Assay for Cell Viability in Pancreatic Cancer

MTT dye reduction assay (Sigma, St. Louis, MO, USA) was carried out to detect the viability of SW1990 and BxPC-3 Cells as previously reported (Dai et al., 2012 (link)). Briefly, cells were seeded into a 96-well plate at a density of 1 × 10⁴ cells/well, cultured for 12 h, then treated with Hellebrigenin with different concentration (0,6, 12, 24, 48, 96 nM in SW1990 and 0, 3.125, 6.25, 12.5, 25, 50 nM in BxPC-3) for 0 to 96 h. At the end of the treatment, 10 μL (50 µg) MTT solution was added into the cells and incubated for another 4 h. 200 μL Dimethylsufloxide (DMSO) was added to each well after removal of the supernatant. After shaking for 10 min, cell viability was measured at the absorbance of 490 nm using an Enzyme-labeling instrument (Multiskan. Go, Thermo Scientific, Ratastie, Finland). The experiments were repeated for three times. Cell growth curve was completed using time as the abscissa and a value (mean ± SEM) as the ordinate.