ScCAD5 cells release more exosomes and PrPSc compared with ScN2a cells, which is inversely correlated to autophagy competence. Comparable numbers of ScN2a and ScCAD5 cells were used in this experiment. A and B, cell lysate and exosomes isolated from conditioned media from ScCAD5 and ScN2a cells were analyzed in immunoblot for HSC70 and PrP. C and D, densitometric analysis for either total PrP or PrPSc respectively from ScCAD5 or ScN2a cell lysate normalized with actin (± S.D.; n = 3 experiments). E, densitometric analysis for exosomal HSC70 normalized with actin in the corresponding cell lysate (± S.D.; n = 3 experiments). *, p < 0.05. F and G, densitometric analysis for either total PrP or PrPSc, respectively, from ScCAD5 or ScN2a exosomes normalized with actin in the corresponding cell lysate (± S.D.; n = 3 experiments). *, p < 0.05. H, immunoblot comparing PrPC levels between CAD5 and N2a cells using mAb 4H11. I, immunoblot comparing the level of LC3-I to LC3-II between ScN2a and ScCAD5 cells with and without treatment with bafilomycin A1 (BA1) for 4 h. Actin was used as a loading control. J, densitometric analysis for LC3-II protein levels normalized with actin (± S.D.; n = 4 replicates). *, p < 0.05, **, p < 0.01.