Trypan Blue Assay for AFB1 Cytotoxicity

To measure cell viability after AFB₁ exposure, we performed a trypan blue exclusion assay. Selected strains expressing CYP1A2 were inoculated in SC-URA until cultures reached an A₆₀₀ ∼0.1-0.5, and then exposed to either 50 μM in AFB₁ or DMSO (solvent) alone. After incubating for 3 hs, cells were washed twice in sterile phosphate buffered saline (PBS) and stained with trypan blue at a final concentration ∼10 μg/ml (Liesche et al. 2015 (link)). Cells were counted in a Nexcelom cellometer T4, according to the manufacturer’s instructions. A minimum of 10⁴ cells were counted and all strains were tested at least twice. Statistical significance was determined by the Student’s t-test.