Figure 6
From: Low doses of bioherbicide favour prion aggregation and propagation in vivo
A6 induces in vitro soluble and/or insoluble rSDS-PrPSc oligomers in a dose dependent manner. (a) Chemical structures of MR100 and A6 compounds. (b) Representative images of yellow precipitates after Rapid Centrifugation Assay (RCA)26. Prion-infected brain samples (50 μL) were incubated with 1.5 mM of MR100 compound (positive control), or with an equivalent volume of DMSO (150 μL, negative control), or with a range of concentrations of A6 compound (from 0.25 mM to 1.5 mM) according to RCA protocol described previously and performed without PK digestion (PK-). After a centrifugation step, a visible precipitate appeared for MR100 (orange colour) and A6 (yellow colour) but not for DMSO sample. The size of the pellet is proportional to the concentration of A6. (c) Western blot analysis of the supernatants and pellets after samples were processed by RCA. Immunoblot was probed with SAF mix antibodies to detect the presence of rSDS-PrPSc. Molecular weight markers are indicated on the left side of the panel. (d) Histogram representing the ratio of the levels of oligomers in the pellet versus oligomers in the supernatant for each concentration of A6. One-way ANOVA (**p-value = 0.0086) followed by Tukey’s multiple comparison test was performed for statistical significance (*p < 0.05, **p < 0.01).
