Figure 3.

AFB₁ enhances intracellular ROS levels and inhibits the transcription of antioxidative stress-associated genes in IMR-32 cells. (A) ROS levels were detected by a carboxy-H2DCFDA probe (Green colour) and observed under a fluorescence microscope. (B) ROS production in IMR-32 cells after AFB₁ exposure was analyzed by flow cytometry after staining with carboxy-H2DCFDA; the data were analyzed using Flow Jo software (BD, Franklin Lakes, USA). (C) The mRNA levels of ROS-related genes in IMR-32 cells were analyzed using qRT-PCR after exposure to 2 μg/mL or 6 μg/mL AFB₁. All the mRNA levels tested were normalized using the mRNA levels of GAPDH as the internal control. All experiments were performed in triplicate and the values represent the mean ± SD of three independent experiments. Statistical significance was defined as * p < 0.05, ** p < 0.01, or *** p < 0.001.