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. 2015 Feb 1;128(3):589–598. doi: 10.1242/jcs.164384

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© 2015. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 5. — InhX-induced NCT ECD movement observed in HEK293 cells using FRET-FLIM. (A) The experimental setup is illustrated. FRET-FLIM was used to monitor the distance between the γ-secretase NCT ECD and NCT CT in HEK293 cells using antibodies carrying donor (green star; rabbit-anti-NCT ECD and donkey anti-rabbit-IgG conjugated to Alexa Fluor 488 as secondary) and acceptor (red star; mouse anti-NCT 9c3 and goat anti-mouse-IgG conjugated to Alexa Fluor 555 as secondary) fluorophores. (B) FRET efficiency was considerably higher following inhX treatment, indicating that the NCT ECD moved closer to the NCT CT on the cytosolic side. Data show the mean±s.e.m.; ***P = 0.0003 (unpaired Student's t-test).