Fig. 2

CORT induces mitochondrial damage in BRL-3A hepatocytes. Hepatocytes were treated with 50 μM CORT for 24 h or 48 h. A-B Mitochondrial cyt c in hepatocytes were determined by western blot (n = 3–4). C, E Immunohistochemical detection of 4-HNE and quantification with ImageJ software to assess lipid peroxidation levels. The region of interest (ROI) of the micrographs was enlarged for clear depiction (n = 3, scale bars: 20 μm). D Hepatocytes were stained with Mito-Tracker Red and 4-HNE, observed with a confocal fluorescence microscope (n = 3, scale bars: 10 μm). F Colocalization analysis was determined by calculating Mander’s coefficient using Coloc 2 in Fiji software (n = 3, 6 cells per experiment). Data were analysed by the Mann–Whitney test or unpaired t-test. ^* P < 0.05, ^** P < 0.01, compared to the vehicle group