Fig. 5.

Effect of rapamycin on TD-induced autophagy and cell death. SH-SY5Y cells were treated with pyrithiamine (0 or 300 nmol/L) and rapamycin (Rapa, 0 or 10 nM) for 5 days. (A) The expression of LC3, p-P70, and t-P70 was determined by immunoblotting. (B) The relative amount of LC3-II was quantified and normalized to GAPDH. p-P70 was quantified and normalized to t-P70. (C) Rapamycin increases TD-induced upregulation of GFP-LC3 puncta in SH-SY5Y cells. Scale bar = 5 µm. (D) The GFP-LC3 puncta per cell were quantified. (E and F) Rapamycin protected TD-induced cell death measured by the cell viability (E) and Annexin-V/PI staining (F). All the results were means ± SEM of three replicates (*p < 0.05, compared with Ct without rapamycin; #p < 0.05, compared with TD without rapamycin).