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. 2017 Mar 10;12:25. doi: 10.1186/s13024-017-0165-0

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© The Author(s). 2017

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 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.

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Fig. 3 — APP-CTF accumulates in endosomal compartments in TSPAN6-overexpressing cells. a Immunofluorescence microscopy of HEK293 cells overexpressing myc-tagged TSPAN6, fixed and probed with a rat monoclonal antibody against myc, a rabbit monoclonal antibody against APP-CTF (Y188), and mouse monoclonal antibodies against EEA1 and LAMP1 to visualize early and late endosomal compartments, respectively. Cells overexpressing myc-TSPAN6 show more APP co-localizing with EEA1 and LAMP1 (filled arrowheads) compared to non-overexpressing cells (empty arrowheads). b Quantification of the number of cells non-overexpressing or overexpressing TSPAN6 showing co-localization of APP-CTF with early (EEA1) or late endosomal (LAMP1) compartments. c Immunoisolation of LAMP1-containing compartments as described [33], after overexpressing Flag-tagged LAMP1 in non-overexpressing or overexpressing myc-tagged TSPAN6. The total inputs and the immunoisolated compartments were analyzed by western blot with specific antibodies against APP, APP-CTF, TSPAN6 and LAMP1. Statistical significance was determined by t-test (***p <0.001)