Fig. 5.
Effects of IND24 treatment on ME7 prion infection. (A) PK-resistant PrP from the brains of terminal animals infected with ME7 prions and treated with vehicle (red) or IND24 (green), as indicated. PrPSc was probed using HRP-conjugated HuM-P Fab. Molecular weight markers of migrated protein standards represent 30 and 20 kDa. This color scheme applies to B, C, and H. (B) Average ratio of monoglycosylated to diglycosylated PrPSc in ME7-infected Tg(Gfap-luc)/FVB mice treated with vehicle (n = 4) or IND24 (n = 5). Error bars represent SD. n.s. = not significant. (C) Conformational stability of ME7 prions from IND24-treated (n = 4) and vehicle-treated (n = 4) Tg(Gfap-luc)/FVB mice. Samples were exposed to increasing concentrations of GdnHCl (0–4 M, as indicated), then subjected to PK digestion. Points represent average percentage of PK-resistant PrP remaining at each concentration of GdnHCl, and error bars represent SD. (D–G) PK-resistant PrP in CAD5 cells (D and E) and LD9 cells (F and G) infected with ME7 (D and F) or ME7[IND24] (E and G), and then treated with 0–20 µM of IND24, as indicated. PK-resistant PrP was assayed by immunoblot using HRP-conjugated HuM-D13 Fab. Images are representative of three replicates. Molecular weight markers of migrated protein standards represent 30 and 20 kDa. (H) Mean survival of mice infected with RML or ME7, then treated with vehicle and/or IND24, initiated at the timepoints indicated. Number of animals per experiment are shown. Error bars represent the SEM. ***P < 0.001, n.s. = not significant, Log-rank test.