Cell viability assay using silibinin and mycotoxin

The cells were counted by Cellometer Auto T4 (Nexcelom Bioscience, Lawrence, MA) and the cell suspension containing cell density 10⁵ cells/mL was split into the 96-well plate. The plates were then incubated for 24 h at 37 °C in humidified atmosphere of 5% CO₂. Then, the tested compounds were added. To assess the effect of silibinin, cells were pre-treated with silibinin at the given concentrations 2 h prior to mycotoxin exposure. After 72 h incubation, the cell viability was tested by standard resazurin assay [114 (link)]. Briefly, the cells were washed three times with 100 µL of PBS and incubated with 100 μL of resazurin solution (0.025 mg/mL) for 3 h. Finally, the fluorescence was measured by a SpectraMax i3x microplate reader (Molecular Devices, UK) at a wavelength of 560 nm excitation/590 nm emission.