Induction of autophagy protects against GCDC-induced cell apoptosis. (A) L02 hepatocytes cells were transiently transfected with GFP-LC3 before treated with GCDC (200 µM), and then administered in the absence or presence of CQ (10 µM) or Rapamycin (2 µM) for 24 hours. (B) The percentage of cells with punctuate GFP-LC3 was measured; at least 30 cells were counted in each individual experiment. Data of at least three replicates were shown as (means ± SD) (*: P < 0.05). (C) In the absence or presence of CQ (10 µM), immunoblots were used to analyze endogenous LC3 expression, with Î²-actin as a loading control. (D and E) L02 hepatocytes were treated as indicated and the apoptosis was analyzed by flow cytometry using Annexin V-FITC. Data were shown individually as the (means ± SD) (*: P < 0.05, **: P < 0.01).