Figure 1
From: Low doses of bioherbicide favour prion aggregation and propagation in vivo
A6 promotes PK rSDS-PrPSc oligomers and interacts with PrP fibrils. (a) Comparison of several compounds for their ability to induce PK rSDS-PrPSc oligomers. Prion-infected N2a58/22L cellular lysates were incubated with 0.5 mM of P30, A6, MR100, α-Ter and A51 for 1 h. Samples were then PK digested at 37 °C for 1 h. Immunoblot was probed with SAF mix antibodies (mixture of three monoclonal anti-PrP antibodies: SAF60, SAF69 and SAF70) for prion detection. Molecular weight markers are indicated on the left side of the immunoblot. The cropped blot is used in this figure and the full-length blot is presented in Supplementary Figure S6. Chemical structures of A6 and α-Ter, 2 compounds described for their herbicidal properties. (b) Fluorescence interaction studies between A6 compound and PrP. Purified full-length recombinant mouse PrP (MoPrP23-230) protein, at 4.4 μM, either soluble or fibrillar, were incubated with 50 μM of A6 compound in 1% DMSO, 50 mM MES pH 6, during 2 h at 25 °C. Emission spectra were recorded between 400 and 550 nm by exciting at λex = 372 nm: 50 μM of A6 (black), 50 μM of A6 + α-soluble MoPrP23-230 (red) and 50 μM of A6 + fibrils of MoPrP23-230 (green). (c) Interaction studies of A6 compound with hamster PrP fibrils. Hamster-S or -R fibrils at a concentration of 4.4 μM were incubated with 40 μM of A6 compound in 1% DMSO, 50 mM MES pH 6, during 2 h at room temperature. Fluorescence spectra were recorded between 400 and 600 nm: 40 μM of A6 (black), 40 μM of A6 + S-fibrils (green) and 40 μM of A6 + R-fibrils (red).
