Fig. 8

Reversing ST2-104 peptide-mediated inhibition of autophagy exacerbates apoptotic cell death in SH-SY5Y cells. SH-SHY5Y cells were treated with 20 mM glutamate plus 20 mM glycine or control medium or with ST2-104 peptide for 24 h at 37 °C and then apoptosis and autophagy levels and apoptosis- and autophagy-related proteins were assessed. a Representative images of autophagosomes. Arrows indicate autophagosomes marked by MDC staining; scale bar, 50 μm. For each well, at least 5 different fields were examined – a representative is shown here. b Apoptosis level was evaluated using the Hoechst 33,258 staining. Scale bar: 50 μm. c, d Detection of LC3-II, Beclin-1, Bax, Bcl-2 and C-caspase-3 protein expression levels using Western blot analysis. Representative blots are shown. Levels of β-actin protein were used as the loading control. Bar represents mean ± SEM from 3 separate wells. ^**P < 0.01, vs. Con group; ^#P < 0.05, ^##P < 0.01, vs. Glu group; ^$P < 0.05, ^$$P < 0.01, vs. Glu + ST2-104 peptide group with one-way ANOVA with Tukey’s post-hoc test