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. 2022 Feb 9;12:2190. doi: 10.1038/s41598-022-06198-4

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© The Author(s) 2022

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 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/.

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Composition of primary bank vole glia cells at the time of infection (20 days post isolation, passage 2). Characterisation was performed by flow cytometry analysis based on expression of GFAP and GLT-1, two specific markers for cells of astrocytic phenotype. Additionally, levels of PrP^C expression were determined as crucial factor for prion propagation. (A) Proportion of the analysed cells, which were found to co-express GFAP and GLT-1. Cell debris and doublets were excluded from the analysis by gating as illustrated. (B) Histogram of those cells, which were stained for GLT-1 and (C) for GFAP. PrP^C expression of each subpopulation is shown in the respective second panel. Red areas of the histograms refer to stained samples and grey areas to the corresponding control incubated without the target antibody. The experiment was carried out in triplicate, of which one representative is shown. According to the analysis the glia cell culture mainly consisted of astrocytes with high PrP^C expression, which provided the basis for an infectible cell assay. FSC: forward scatter, SSC: side scatter.