The Intracellular ROS levels were detected by DCFH-DA (Beyotime Biotechnology, S0033S). The Intracellular mitochondrial ROS levels were detected by Dihydrorhodamine 123 (MCE, HY-101894). Cells were cultured in six-well plates at a density of 2.0 × 105 cells/well. After being treated with l -sorbose, the cells were gently washed with D-PBS (BBI, E607009) followed by incubation with DCFH-DA or Dihydrorhodamine 123 at 37 °C for 30 min.
To obtain microscopy images, cells were seeded on a confocal dish overnight and treated withl -sorbose for 6 h. After being treated with l -sorbose, the cells were gently washed with D-PBS followed by incubation with MitoBright LT (Dojindo Laboratories, MT11) and DCFH-DA (Dojindo Laboratories, CK04) at 37 °C for 30 min, or Dihydrorhodamine 123 alone. Then the cells were washed with D-PBS twice. Images were obtained with a Nikon C2 Eclipse Ti-E inverted confocal microscope equipped with NIS-Element AR software.
To measure the volume of mitochondria, cells were cultured in six-well plates at a density of 2.0 × 105 cells/well. After being treated withl -sorbose, the cells were gently washed with D-PBS followed by incubation with MitoBright at 37 °C for 30 min. Then the cells were washed twice with D-PBS. The fluorescence of the cells was measured immediately on a FACSCalibur flow cytometer and analyzed by FlowJo software.
To obtain microscopy images, cells were seeded on a confocal dish overnight and treated with
To measure the volume of mitochondria, cells were cultured in six-well plates at a density of 2.0 × 105 cells/well. After being treated with