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. 2019 Oct 10;9:14656. doi: 10.1038/s41598-019-51084-1

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Detection of PrP^Sc in the brain, spleen and granuloma of L-BSE or C-BSE-infected mice. At various times post-infection, groups of animals were sacrificed and brain, spleen and granuloma were removed for tissue processing and analysis. (A) proteinase-K digestion followed by western blot analysis using biotinylated Sha31 anti-PrP antibody. Note that the exposure times for brain and spleen WB membranes ranged from 3 to 5 minutes, while the granuloma samples required longer exposure times (over 3 hours). (B) Immunofluorescence (IF) detection of PrP^Sc in the spleen or granuloma of tg338 mice inoculated with L- and C-BSE prions. Labeling of PrP^Sc in spleens (B-1, B-3, B-5) and granulomas lymphofollicular structures (B-2, B-4, B-6) from control (B-1, B-2), L-BSE-infected animals (B-3, B-4) or C-BSE-infected animals (B-5, B-6); (C) PMCA and RT-QuIC templating activity of PrP^Sc present in individual brains, black ■; spleens, red ●; and granulomas, blue ▲, from C-BSE or L-BSE-inoculated mice. Gray squares represent the inoculum titer. Open symbols represent PrP^0/0 animals. See Supplementary Table 1 for a percentage of positive samples.