Fig. 5. FRAP analysis showing that there is active exchange between caveolin-1-GFP located in the peri-centrosomal region and the other pools of caveolin. Cells stably expressing caveolin-1-GFP were plated 48 hours before viewing into culture dishes that had a glass coverslip glued to the bottom. Confocal images were taken at 1.6 second intervals for approximately 1 minute and then the entire peri-centrosomal region of the upper cell was bleached in 6 spots for 2 seconds each using the 488 laser at full power. Immediately following the bleach, images were again collected at 1.6 second intervals for another 5 minutes and 40 seconds. Panel A shows the prebleach image and peri-centrosomal accumulations of caveolin-1-GFP in two cells. The central accumulation in the top cell was then bleached (area within the white circle, B). Panel C shows that the accumulation at the center of the cell has recovered within 5 minutes, indicating that caveolin-1-GFP travels between the various caveolin positive compartments. The corresponding movie can be viewed online (http://jcs.biologists.org/supplemental).

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