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Figure 2


Fig. 2. PKC activity drives the interaction of fascin with PKC{gamma}. (A) SW480-Pa cells transiently expressing GFP-fascin, alone or plus mRFP-PKC{gamma}, were treated with solvent or PDBu for the indicated times, and then fixed and imaged using FLIM to measure FRET. PDBu and TPA activate conventional PKC equivalently at early timepoints. (B) Quantification of data from nine cells from three independent experiments as in A. (C) SW480-Pa cells transiently co-expressing either GFP-fascin-S39A or GFP-fascin-S39D with PKC{gamma}-mRFP were treated with PDBu for 20 minutes, then fixed and imaged using FLIM to measure FRET. Only GFP-fascin-S39D interacted with active PKC{gamma}. (A,C) Intensity multiphoton GFP images (donor) and, where relevant, epifluorescence images for RFP (acceptor). Lifetime images are depicted using a pseudocolour scale where red is a low lifetime (1.8 nseconds) and blue is high (2.3 nseconds).