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Figure 6


Fig. 6. FRAP analyses reveal two pools of wt GAD65-GFP replenishing the Golgi in primary rat hippocampal neurons. The Golgi-associated pool of palmitoylation-deficient (A) and wt GAD65-GFP (B) in living rat hippocampal neurons at DIV10 were selectively photobleached (t=0) and fluorescence recovery was monitored over time at 37°C. (C) Kinetics of fluorescence recovery after bleaching of the Golgi-associated pool of wt GAD65-GFP (red circles, n=10 neurons) and GAD65(C30,45A)-GFP (blue circles, n=10 neurons). Recovery of GFP fluorescence into the photobleached Golgi compartment was recorded at low laser power and normalized taking into consideration the loss of GFP fluorescence in the cell during the recording. Data are mean ± s.d. for a representative experiment. (D) Half-times of Golgi fluorescence recovery following photobleaching for wild-type and palmitoylation-deficient GAD65-GFP. (E) Percentage recovery of Golgi fluorescence following photobleaching. Data for D and E are the mean ± s.e. from five independent experiments (5-10 cells per experiment).





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