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Video 1 (6 MB)
VSVG-GFP in^{the Golgi complex (FRAP). COS7 cellswere transfectedwith VSVG-GFP, placedonglass-bottom microwell disheswith coordinatedgrids, subjectedto small-pulse protocol, andexaminedby laser scanning confocal microscope.4 minafter the shiftfrom 15°C to 40°C, the Golgi spotscontaining VSVG-GFP inthe central Golgi area were maskedby a high-ER background.To remove thisbackground, the whole cell, exceptthe Golgi area, wasrepeatedly bleached.Thismade the fluorescentspotsinthe Golgi zone more evident.These spotshoveredwithinthe Golgi area withoutexhibiting long-range movements.Next, at6-7 min, half of the Golgi area wasbleachedto examine whether fluorescence wouldrecover inthe bleachedspots.No recovery wasobserved.10 minafter the shiftfrom 15°C to 40°C, the GFP-containing spotsbecame more mobile andbeganto leave the Golgi area.}