The tight-binding AChEis huprine Y and Hup8TH efficiently decrease PrP
accumulation in Scrapie-infected MovS6 cells. a Scrapie-infected MovS6 cells were incubated with 20 μM of the indicated AChEis for 6 days and then lysed. PrPSc accumulation was determined by using the anti-PrP antibody Sha31b; the anti-β-actin antibody was used to control protein loading. Ctr, vehicle-treated cells. b Prion-infected MovS6 cells were incubated with 0.5 μM Hup8TH or 0.1 to 0.5 μM Hup8TH and then processed as in a. c Densitometric measurement of PrPSc signal, expressed as percentage relative to the untreated control (closed circles). Results are expressed as the mean ± SD of three different experiments. Cell viability determined by using the MTT assay is also shown (open circles). The absorbance values are expressed as the mean ± SD of four independent experiments relative to the absorbance values of the untreated control. Solid and dashed lines are fits to the data. d Protein lysates from uninfected MovS6 cells were analyzed by immunoblotting without proteinase K digestion. The effect of Hup8TH on PrPC accumulation was determined using the SAF32 antibody. Molecular weights (in kilodaltons) are indicated on the left of the blots.