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Fig. 2. FLIM analysis of the proximity of wild-type BACE and GGA1 within cells. N2a cells were transfected with BACE-V5 (labeled with FITC) and GGA1-Myc (labeled with Cy3). Only the donor fluorophore (FITC) was applied for the negative control (A,B). The intensity image shows the standard immunostaining pattern for BACE (A,C). The color-coded FLIM image shows the lifetimes (in picoseconds) of FITC in the absence (B) or presence (D) of the acceptor Cy3. The shorter FITC lifetimes reflecting proximity between GGA1 and BACE appear only in juxtanuclear compartments (TGN) (D). The negative control exhibits a single population of lifetimes for FITC in the absence of an acceptor, represented by a homogenous blue-green pseudocolor image.





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