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. 2017 Jan 31;10:15. doi: 10.3389/fnmol.2017.00015

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Copyright © 2017 Canu, Pagano, La Rosa, Pellegrino, Ciotti, Mercanti, Moretti, Sposato, Triaca, Petrella, Maruyama, Levi and Calissano.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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TrkA and APP interaction in septal primary neurons measured by proximity ligation assay. (A) Confocal microscopy analysis of double-staining of APP (rabbit APP-CT A8717, red) and Trk (mouse Trk B3, green) in primary septal neurons showing co-localization of APP and TrkA. (B) The TrkA/APP complex was visualized by PLA, which generates red dots when the two proteins are in close proximity. Mouse and rabbit anti TrkA-CT (mouse Trk B3 and rabbit TrkA ab7261) and anti APP-CT (rabbit APP-CT A8717 and mouse APP clone C1-6.1) were used as primary antibodies and anti-mouse MINUS and anti-rabbit PLUS were used as secondary antibodies. Red fluorescent dot represents single interaction between TrkA and APP. (C) In addition to PLA (red dots), rat primary septal neurons were immunostained with goat anti ChAt (green) and with DAPI for nuclei (blue). (D) PLA assay using mouse anti APP (C1-6.1) and rabbit anti TrkB (sc-119) or TrkC (sc-117).