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. 2020 Jun 19;8:85. doi: 10.1186/s40478-020-00966-x

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© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 2 — Immunoblot and N-terminus epitope mapping of resPrP^D from sCJDMV2 variants, sCJDMV1 and controls. a both resPrP^D types 1 and 2 (arrows) from all MV variants reacted with the type-non-specific Ab 3F4. The three resPrP^D major glycoform are present in all preparations. The unglycosylated components shows the ~ 21 kDa mobility of resPrP^D type 1 in -MM1 and -MV1 (dotted arrow), and the 19 kDa mobility of resPrP^D type 2 (solid arrow) in all other preparations. Both “pure” sCJDMV2K and the mixed variant MV2K-C (50:50%) exclusively display the 20 kDa band (dashed arrow) along with the 19 kDa type 2 component (solid arrow). b type 1-specific Ab 12B2, besides resPrP^D type 1 of MM1 and MV1, also demonstrated all three glycoforms of the 20 kDa component in sCJDMV2K and MV2K-C (dashed arrow) confirming the type 1-like N-terminus immunoreactivity of the 20 kDa as well as of its other two glycoform components; note that unlike with 3F4, when probing with Ab 12B2 the “20” kDa band (dashed arrow) aligns with the unglycosylated isoform of MM1 and MV1 at 21 kDa. c Ab Tohoku-2, type 2-specific, reacted only with all resPrP^D type 2 as indicated by the 19 kDa unglycosylated isoform (solid arrow). d SAF32 (to slightly more N-terminal epitopes than 12B2) recognizes resPrP^D type 1 of MM1 and MV1 but not (or very weakly) resPrP^D associated with MV2K and MV2K-C. All tissue samples were obtained from the frontal cortex. ^* Residue 97 (or another C-terminally closely adjacent N-terminus) is required for Tohoku-2 to immunoreact [17]