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. 2017 Dec 14;7:17565. doi: 10.1038/s41598-017-17770-8

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© The Author(s) 2017

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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Short-term treatment with AR-12 reduces PrP^Sc levels in three persistently prion-infected cell lines. (a) Persistently prion-infected neuroblastoma cells (ScN2a; prion strain 22 L) were treated for 72 h with 0.5, 1, 1.5, 2, 2.5 or 3 µM of AR-12. Solvent only-treated cells (DMSO) were used as control. Cells were lysed and lysates split into two halves. One was treated with proteinase K (PK; 20 µg/ml for 30 min at 37 °C) and both subjected to immunoblot analysis. Immunoblot was developed with anti-PrP monoclonal antibody (mAb) 4H11 and the blot was re-probed with a mAb for actin (gel loading control). (b) Densitometric analysis of ScN2a immunoblots. Data are represented as a percentage of vehicle-treated (DMSO) cells from at least three independent experiments. (c) Persistently prion-infected neuronal ScCAD5 cells (prion strain 22 L) were treated for 72 h with 1, 2, 3, 4 or 5 µM of AR-12. Solvent only-treated cells (DMSO) were used as control. Immunoblot was developed with anti-PrP mAb 4H11 and the blot was re-probed for actin (gel loading). PrP^Sc was reduced after AR-12 treatment. (d) Densitometric analysis for ScCAD5 immunoblots. Data are represented as a percentage of vehicle-treated (DMSO) cells. (e) Persistently prion-infected mouse embryonic fibroblasts (MEF) (prion strain 22 L) were treated for 72 h with 0.5, 1,1.5, 2, 2.5 or 3 µM of AR-12 and analyzed as above. (f) Densitometric analysis for ScMEF immunoblots. Data are represented as a percentage of vehicle-treated (DMSO) cells. (*p < 0.05), (**p < 0.01) (***p < 0.001) considered significant.