Figure 2.

Galantamine inhibited the amyloid beta‐peptide (Aβ)‐induced cytostatic autophagy. (A) PC12 cells incubated with Aβ (2, 5 μmol/L) for 24 h were analysed by western blotting for LC3, BECLIN‐1, p62 and β‐actin. The effect of autophagy inhibitor 3‐MA (5 mmol/L) and CQ (10 μmol/L) on expressions of LC3, BECLIN‐1, p62 (B and C), caspase‐3 and cleaved‐caspase3 (D and E) were detected in PC12 cells treated with Aβ (2 μmol/L) for 24 h. (F) PC12 cells were treated with Aβ (2 μmol/L) for 24 h after pretreatment with 3‐MA (5 mmol/L) for 2 h or with CQ (10 μmol/L) for 6 h. Cell apoptosis rate was measured by TUNEL assay, and then, (G) cell proliferation was examined by the MTT assay. (H) PC12 cells were transfected with siNC or siATG5 for 24 h, and then treated with 2 μmol/L Aβ for another 24 h. The expression of ATG5, LC3, Beclin 1, p62 and β‐actin were examined by western blotting, (I) Cell apoptosis rate was measured by TUNEL assay and (J) cell viability was examined by the MTT assay. (K) The cells treated with 2 μmol/L Aβ or galantamine (40 μmol/L) for 24 h, then immunofluorescence assay was used to determine LC3 accumulation, and (L) western blotting was employed to assess the expression of LC3, BECLIN‐1, p62. β‐actin was used as a loading control. The results of western blotting were analysed using ImageJ (mean ± SD of 3 independent experiments)