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. 2023 Jun 1;11:87. doi: 10.1186/s40478-023-01577-y

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© The Author(s) 2023

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/. 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. 3 — Presence of APP WT and APP FAD mutants at the cell surface. A Cell surface staining. HeLa cells were transiently transfected with N-terminally c-myc-tagged APP WT, Swedish, Arctic, Flemish, Iowa or Iberian. Cells were labeled with α-c-myc antibody (green, surface APP; red, total APP). The images were taken with an epifluorescence microscope including apotome. Scale bar: 20 μm. B ICC quantification. The relative surface APP intensity [%] indicates the percentage of APP present at the surface compared to total APP intensity. Bars represent mean values ± SEM; n = 18 (three biological replicates). Statistical analysis using one way ANOVA with Tukey`s post hoc test did not reveal any significant differences. C Cell surface biotinylation of transiently APP WT, Swedish, Arctic, Flemish, Iowa or Iberian transfected HEK293T cells. Direct load of cell lysates is shown in the left panel. In the right panel, APP cell surface levels after streptavidin immunoprecipitation and Western blot detection with anti-c-myc antibody are shown. β-Actin served as a loading control and was detected in the IP samples only to a minor extent, which confirms specificity of the pull down of surface proteins. The lower panel shows a higher exposure time