Surface delivery of CAV1-mEGFP in U18666A-treated cells. CV1 cells treated with 5 µM U18666A (16 h) were transfected with CAV1-mEGFP using electroporation (see Materials and methods) and incubated in the presence of 1 mM cycloheximide during cell attachment (2 h). Expression of CAV1-mEGFP was induced by washout of cycloheximide, and cells were imaged by TIR-FM 60 min after washout. The evanescent field was adjusted such that both cell surface and part of the Golgi were visible. CAV1-mEGFP can be observed in tubular post-Golgi carriers. Upon arrival at the cell surface, they release their cargo, visible as burst of fluorescence diffusing laterally into the plasma membrane. Boxed areas showing examples are enlarged below. Time-lapse sequences were acquired at 1 Hz with an integration time of 500 ms/frame. Video is displayed at 15× real time.
