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Figure 2


Fig. 2. Exo70 interacts with activated TC10 in PC12 cells. (A-O) mCFP-labeled wild-type (wt) TC10{alpha} (A,C,F,H) or dominant-negative TC10{alpha}-T23N (K,M) were expressed alone (A,F,K) or co-expressed (C,H,M) with mVenus-Exo70 (D,I,N). mCFP fluorescence lifetimes ({tau}) (B,E,G,J,L,O) were imaged by two-photon-time domain FLIM. Lifetimes are indicated in false colour ranging from 1.5 nanoseconds (blue) to 2.5 nanoseconds (red). Reduced lifetimes can be detected for mCFP-TC10{alpha} interacting with mVenus-Exo70 upon NGF-induction (E, n=15), but not in non-treated cells (J, n=12) or for mCFP-TC10{alpha}-T23N upon co-expression with mVenus-Exo70 (O, n=8). Bars, 5 µm (untreated cells), 10 µm (NGF-treated cells). (P) Shown are cumulative histograms of cellular lifetime distributions for FRET between mCFP-TC10{alpha} forms and mVenus-Exo70. Letters at the traces refer to the respective indicated conditions in the above panel (E,J,O). Note the lower lifetimes (higher FRET efficiencies) for NGF-treated mCFP-TC10{alpha}-co-expressing PC12 cells only (E).





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