Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

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

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

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.

PMC Copyright notice

Visualization of TrkA-APP protein interaction using FRET and BiFC analysis. (A) Confocal microscope images of FRET acceptor photobleaching assay of HEK293 cells co-transfected with APP-CFP and TrkA-YFP. The CFP and YFP images before and after YFP photobleaching, the merged image of CFP and YFP prior to photobleaching, and pseudo-colored image showing an FRET efficiency values map. (B) The schematic diagram of dimerization pairs used in the BiFC analysis. (C) HEK293 cells were transfected with TrkA-VN alone, TrkA-VC and ERB-VN, APP-VN alone, TrkA-VC and TrkA-VN, TrkA-VC and APP-VN. Fluorescence images of HEK293 cells expressing the proteins indicated in each panel were acquired by confocal microscopy 18–24 h after transfection. (D) Four images of cells transfected with TrkA-VC/APP-VN were selected to show cells with different level of protein expression (a, b: high level of expression; c, d: low level of expression) but similar pattern of subcellular distribution; e, APP staining with mab 22C11 (red). (E) BiFC analysis shows that TrkA-VC interacts also with APP-Sw/In-VN but not with APP-Y682G-VN, despite this latter is efficiently expressed when transfected (see immunostaining for APP performed with mab 22C11, red). (F) Cell lysates of HEK293 co-transfected with TrkA and APP-Sw/In or APP-Y693G were immunoprecipitated with mab Trk (B3) antibody or mab Bcl2 (IgG) as control followed by immunoblot with rabbit APP-N-terminal and rabbit TrkA.^∗ and ^∗∗ indicate the IgG heavy and light chains respectively.