Glycochenodeoxycholic acid induces autophagy in L02 hepatocytes. (A and B) The L02 normal hepatocytes cells were transfected with GFP-tagged LC3 and then pretreated with GCDC (200 µM) as indicated (for 2, 12, and 24 hours). Images were taken under a fluorescence microscopy. GFP-LC3 puncta (mean ± SD) were quantified for each experiment. At least 20 cells were counted in each individual experiment. (**: P < 0.01). (C) Electron micrographs showing the cells ultrastructure as indicated. Black arrows indicate the autophagic vacuoles in the cytoplasm. (Bar: 20 µm). (D) The number of autophagosomes were quantified (mean ± SD) (*: P < 0.05) (E) Whole cell lysates were subjected to western blot to detect the expression of LC3 and p62 as indicated (for 0, 6, 12 and 24 hours). (F) LC3 and the p62 as indicated (for 25, 50, 100, 200, 400 µM) were examined by immunoblotting.