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. 2020 Jan 28;10:1358. doi: 10.1038/s41598-020-58476-8

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© The Author(s) 2020

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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Rab5^DN mutant triggers the interaction between APP and BACE1. For confocal analyses, N2a cells were transfected with APP:VN and BACE1:VC expressing vectors. Only where indicated, cells additionally expressed mCherry-Rab5^DN (Rab5^DN), mCherry-Rab5^CA (Rab5^CA), or underwent vardenafil treatment (VDF). Nuclei are blue-fluorescent, the green fluorescence is indicative of APP-BACE1 interaction, whereas the red fluorescence is due to the expression of mutant mCherry-Rab5. Note the increase in the APP-BACE interaction in cells treated with vardenafil compared to control cells and how this effect is mimicked by Rab5^DN but not by Rab5^CA expression. The inset shows merged blue/green fluorescence, and mCherryRab5 expression, at a greater magnification. White scale bar = 50 μm; black scale bar = 20 μm. Figure is representative of 3 independent experiments, all showing essentially similar results.