Effects of PI3K, MEK and lysosomal inhibitors on rapamycin-elicited autophagic responses in N2a-FK cells.
(A) N2a-FK cells were co-treated with LY294002 (10 μM) or PD98059 (50 μM) along with rapamycin (1 μM) for 48 h. (B) N2a-FK cells were co-treated with NH4Cl (10 mM) together with rapamycin (1 μM) for 48 h. Upper: PK-treated or-untreated samples were applied at concentrations of 100 and 50 μg protein per lane onto a 15% polyacrylamide gel and subjected to SDS-PAGE. The proteins were detected by western blotting using anti-PrP,-LC3 and -β-actin antibodies. Bottom: PrPSc expression levels following drug treatment were quantified in N2a-FK cells. The results are representative of at least three independent experiments, with each experiment performed in triplicate. *p < 0.05 and **p < 0.01 (one-way ANOVA followed by Tukey's test).