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Fig. 2. Suramin and derivatives reduce the amount of PrP^Sc in prion-infected cells to undetectable levels and induce insoluble full-length PrP aggregates. (A,B,C) The effect of suramin derivatives on PrP^Sc was determined in prion-infected neuroblastoma cells (3F4-ScN2a) using a solubility assay and proteinase K (PK) treatment followed by immunoblot analysis. Cells were seeded on 60 mm plates and treated daily with suramin or with one of its derivatives, as indicated. All compounds were applied at a concentration of 200 µg/ml for 4 days. Mock-treated cells were used as a control. Cells were harvested and postnuclear lysates were either treated with 20 µg/ml PK for 30 minutes at 37°C or subjected to ultracentrifugation at 100,000 g in the presence of 1% sarcosyl, to separate soluble (S, supernatant) from insoluble (P, pellet) fractions, as indicated above the blots. PrP was visualised by immunoblotting using the monoclonal anti-PrP antibody 4H11. Molecular size markers are depicted on the left. No PK-resistant PrP is detected after treatment of the cells with NF058, NF078 (A, lanes 10,16); NF110, NF305, NF307, NF506 (B, lanes 1,4,7,10); suramin, NF023 or NF449 (C, lanes 4,7,10). NF542 and NF710 were highly cytotoxic. Actin loading controls are shown below each panel.

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