Fig. 2
Steroid intermediates stimulate reactive oxygen species (ROS) production and oxidative stress in Y1-BS1 cells. Y1-BS1 cells were pre-loaded with 10 μM DCFH-DA for 30 min, washed and incubated with HDL3 (30 μg/ml), 22(R)-diol (10 μM) or pregnenolone (10 μM) for different time point. At the end of incubation, the media were removed and cells were immediately lysed and centrifuged. DCFH-DA fluorescence of the cell lysates was measured using a microplate reader with excitation and emission wavelengths of 480 and 525 nm respectively. Results are summary of three independent experiments with triplicates in each experiment. * p < 0.05, ** p < 0.01, ***p < 0.001