Figure 2.

CGA inhibits Aβ_25-35-induced autophagy in SH-SY5Y cells.

Notes: (A) The viability of cells treated with various concentrations of CGA for 24 and 48 h was determined with MTT assay. (B) Representative MDC-stained autophagy vesicles in SH-SY5Y cells (right panel, ×200), and analysis of mean fluorescence intensity (left panel). (C) Representative Western blot imagines of LC3B-II/LC3B-I (16/18 KDa) and p62/SQSTM (75 KDa) in SH-SY5Y cells (left panel); β-actin (43 KDa) was used as a control for protein loading. The right panel shows the relative optical density values of LC3B-II/LC3B-I and p62/SQSTM between the experimental groups. (D) Representative Western blot imagines of beclin1 (50 KDa) and Atg5 (32 KDa) in SH-SY5Y cells (left panel); GAPDH (37 KDa) was used as a control for protein loading. The right panel shows the relative optical density values of beclin1 and Atg5 between the experimental groups. (E) SH-SY5Y cells expressing mCherry-GFP-LC3 stably were treated in the presence or absence of CGA. Autophagosomes (yellow foci) and autophagic flux (red-only foci) were measured with confocal fluorescence microscopy (FV1200; Olympus, Tokyo, Japan). Three independent experiments were performed, and the date are expressed the mean ± s.e.m for five samples per treatment group. **P<0.01 compared with the no Aβ_25-35 treatment group; ^#P<0.05, ^##P<0.05, ^###P<0.001 compared with the Aβ_25-35 treatment group.