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. 2009 Dec;134(6):489–521. doi: 10.1085/jgp.200910314

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© 2009 Moss et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jgp.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 6. — FRET assessed by acceptor photobleach. (A) Plots of acceptor photobleach and donor photorecovery for mGAT1CFP*/mGAT1YFP* (n = 10) or (B) mGAT1CFP8/mGAT1YFP8 (n = 7) coexpressed in N2a cells. Data for each fluorophore were normalized to absolute fluorescence at time zero and fitted to a single exponential plus finite baseline: $y = y_{0} + A_{1} e^{(- (x - x_{0} / τ_{1}))}$ . (C) A scatter plot of CFP photorecovery versus YFP photobleach for the FRET pairs in A and B. FRET efficiency for each transfection was calculated by extrapolating to the linear regression fits to YFP photobleach = 1.0. The Y-intersect (I_D) was then entered into Eq. 1. (D) Plot of acceptor photobleach and donor photorecovery for mGAT1CFP45/mGAT1YFP45 (n = 12). (E) Mean acceptor photobleach and donor photorecovery for the CFP15mGAT1/YFP15mGAT1 cotransfection in N2a cells (n = 5).