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50 units of Rocky Mountain Laboratory (RML) scrapie prions. Mice were monitored every second day, and scrapie was diagnosed according to standard clinical criteria (6).
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26
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0343640929
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note
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50 units of mouse-adapted scrapie prions (RML 4.1). Depletion was maintained by weekly administration of 100 μg of the fusion protein for 8 weeks. A second set of mice were inoculated with prions as above; 300 μg of LTβR-lg were injected 1 week later and 100 μg LTβR-lg were injected once a week for a further 6 weeks. Three mice from each group were culled 1, 3, and 8 weeks after inoculation, respectively. As control, C57BL/6 mice were subjected to the same regime with a human IgG polyclonal preparation (hulgG, Sandoglobulin) instead of LTβR-lg.
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27
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data not shown
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Spleen homogenates (10% w/v) from frozen tissues were prepared in 2% sarkosyl-phosphate-buffered saline (PBS). Homogenates were adjusted to 8 mg/ml protein. Total protein (200 μg) of each spleen was treated with proteinase K (20 μg/ml, 30 min, 37°C). Samples were subjected to electrophoresis through an SDS-polyacrylamide gel (12.5%), transferred to nitrocellulose membranes, and probed with polyclonal antiserum 1B3 to mouse PrP (44), and bound antibody was visualized by enhanced chemiluminescence (SuperSignal West Dura Extended Duration Substrate, Pierce).
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0343205304
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note
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We thank M. Kosco for the gift of FDC M1 antibody and C. Musahl for the rabbit anti-PrP antiserum XN. Supported by the Kanton of Zürich and by Schweizerischer Nationalfonds and European Union grants to A.A. and C.W.
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