Figure 4.

Effect of Carnosic acid (CA) and Carnosol (CS) treatment on PrP^Sc levels (in vitro assays). N2a22L cells were incubated with 0.5 × LD₅₀ of CA and CS for 48 h or left untreated (Control), lysed, and then divided into two different fractions. 0.5 × LD₅₀ was estimated at 14.84 μΜ for CA and 16.53 μΜ for CS for N2a58 and at 13.58 μΜ CA and 12.85 μΜ CS for N2a22L. Fraction lysates were either treated (+) or not (-) with PK, and 50 μg of PK-untreated or 100 μg of PK-treated fraction was resolved by SDS-PAGE and immunoblotted with the anti-PrP antibody 6H4 (1:5000) and an anti-β-actin (1:2000) antibody. One representative blot for each compound, along with the densitometric analysis from three independent experiments, are depicted. Data represent means ± SD; stars denote statistical significance (unpaired, one-tailed, T-test); *: p value < 0.05, **: p value < 0.01. CA: dark grey, CS: light grey, Control: black.