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. 2019 Aug 1;9:11151. doi: 10.1038/s41598-019-47629-z

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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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Stable infection of CAD5 cells reconstituted with Mo-PrP and BV-PrP by 22 L prions. (a) Immunoblot analysis demonstrating sustained infection phenotype of reconstituted MoPrP cells. Control CAD5 cells (C) and cells reconstituted with mouse PrP (CAD5_Mo) were infected with 1% brain homogenate from terminal ill mice infected with 22 L prions (CAD5_Mo_22 L). Cell lysates from passage P3 of parental CAD5 and passages P3 and P9 of the CAD5_Mo_22L cells were treated with and without proteinase K. The non-treated (−PK) represents the total PrP (left panel) while the treated lysate (+PK) represents PrP^Sc (right panel). The reconstituted CAD5-Mo cells show presence of PrP^Sc from passage P3 to P9 post-infection, demonstrating successful and persistent infection. (b) Immunoblot analysis demonstrating sustained infection of CAD5 KO cells reconstituted with bank vole PrP (CAD5_BV). Control CAD5 (C) and the CAD5_BV cells were incubated with 1% brain homogenate from mice infected with 22L prions (CAD5_BV_22L) and processed as above. The reconstituted CAD5_BV cells showed presence of PrP^Sc from passage P3 to P9 post-infection (right panel), demonstrating successful and persistent infection.