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Volume 19, Number 12—December 2013

Research

Spontaneous Generation of Infectious Prion Disease in Transgenic Mice

Juan-María TorresComments to Author , Joaquín Castilla, Belén Pintado, Alfonso Gutiérrez-Adan, Olivier Andréoletti, Patricia Aguilar-Calvo, Ana-Isabel Arroba, Beatriz Parra-Arrondo, Isidro Ferrer, Jorge Manzanares, and Juan-Carlos Espinosa
Author affiliations: Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria, Madrid, Spain (J.-M. Torres, J. Castilla, B. Pintado, A. Gutiérrez-Adán P. Aguilar-Calvo, A.-I. Arroba, B. Parra-Arrondo, J.-C. Espinosa); Basque Foundation for Science, Bilbao, Spain (J. Castilla); Ecole Nationale Vétérinaire de Toulouse, Toulouse, France (O. Andréoletti); Hospitalet de Llobregat, Barcelona, Spain (I. Ferrer); Universidad Miguel Hernandez, Sant Joan d´Alacant, Spain (J. Manzanares)

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Figure 3

Host cellular prion protein (PrPC) solubility and proteinase K (PK) resistance studies in homozygous 113LBoPrP-Tg037, 113LBoPrP-Tg009, and control BoPrP-Tg110 mice. Western blot analysis with monoclonal antibody 2A11 of soluble (S) and insoluble (P) fractions obtained from mouse brain extracts (5% sarkosyl in phosphate-buffered saline, pH 7.4, previously cleared by centrifugation at 2,000 × g) after ultracentrifugation at 100,000 × g for 1 h. P fractions were treated with 5 μg/mL of PK (PK+) at

Figure 3. . . Host cellular prion protein (PrPC) solubility and proteinase K (PK) resistance studies in homozygous 113LBoPrP-Tg037, 113LBoPrP-Tg009, and control BoPrP-Tg110 mice. Western blot analysis with monoclonal antibody 2A11 of soluble (S) and insoluble (P) fractions obtained from mouse brain extracts (5% sarkosyl in phosphate-buffered saline, pH 7.4, previously cleared by centrifugation at 2,000 × g) after ultracentrifugation at 100,000 × g for 1 h. P fractions were treated with 5 μg/mL of PK (PK+) at 37°C for 60 min or left untreated (PK–). Panels A and B, show brain extracts from diseased 113LBoPrP-Tg037 mice or from 500-day-old 113LBoPrP-Tg009 and BoPrP-Tg110 mice. Panel C shows brain extracts from 30-day-old mice. In panels A and C, equivalent amounts of brain material were solubilized, centrifuged, and loaded onto the gel. In panel B, an 8-fold (8×) equivalent brain tissue mass was loaded to obtain equivalent PrPC signals for the other 2 mouse lines. 113L, leucine substitution at codon 113; BoPrP, bovine prion protein. Values on the right are molecular masses in kilodaltons.

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