Figure 5.

A-769662 facilitates H₂O₂-induced autophagy activation. MG-63 cells were treated with hydrogen peroxide (H₂O₂, 250 μM), cells were further cultured for indicated time points, and western blots were performed to test LC3B, Beclin-1, p62 and β-actin (A); The expressions of LC3B, Beclin-1, p62 and β-actin in H₂O₂ (250 μM) stimulated MG-63 cells transfected with GFP-shRNA, AMPKα shRNA lentivirus or AMPKα kinase dead (KD) plasmid were tested (B); MG-63 cells were pre-treated with 3-methyladenine (3-MA, 1 mM), or Bafilomycin A1 (BafA1, 0.1 μM) for 1 h, followed by H₂O₂ (250 μM) stimulation for 72 h, cell viability (C) and trypan blue assay (D) were performed; MG-63 cells were treated with H₂O₂ (250 μM), A-769662 (10 μM) or both for 24 h, the expressions of LC3B, Beclin-1, p62 and β-actin were tested (E); The viability of MG-63 or MC3T3-E1 cells with indicated treatment for 72 h was tested by MTT assay (F and G). # p < 0.05.