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Fig. 5. FLIP analysis reveals that GFPSmad4 is more mobile in the cytoplasm than is GFPSmad2. FLIP analysis was performed on either HeLa TK- cells transiently transfected with plasmids expressing GFPSmad2 or GFPSmad4 and FLAG-Smad2 (A) or HaCaT cell lines stably expressing GFPSmad2 or GFPSmad4 (B). Each row shows the fluorescence image prior to bleaching (time=0 seconds) and then after three consecutive 80-second bleaching periods. The fluorescence was quantitated at the bleach point (red diamond) and at a distant reporting point in the cytoplasm of the same cell (blue square) and at a reporting point in the cytoplasm of an adjacent cell (white triangle). In each case the fluorescence was normalised to the initial fluorescence prior to photobleaching, and the relative fluorescence was plotted. The data are representatives from at least three different experiments.
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