Figure 3.

Inhibition of GSK3β further promotes autophagy induced by starvation in vitro. (a). Transfected GFP-LC3 plasmid for 12 h, the primary hepatocytes were pre-incubated with SB216763 (2, 5 or 10 μM) for 12 h to observe the formation of autophagosomes. (b) The primary hepatocytes were stimulated with or without SB216763 for 30 min, followed for 3, 6 or 12 h. The cell lysates were analyzed by western blot in different time. The levels of LC3B, p62, Atg7, Atg5, Beclin-1and β-actin were measured by western blotting. The graph of densitometry analysis should be shown (compared with starvation 3 h group: *P<0.05; compared with starvation 6 h group: ^#P<0.05; compared with starvation 12 h group: ^&P<0.05). (c) Inhibition of GSK3β induced autophagic flux in primary hepatocytes under starvation. Western blot analysis of expression of the LC3B and p62. A representative results from three independent experiments is shown. Densitometry analysis of the proteins was performed for each sample