CIL:24067

E-cadherin local dynamics were studied in mature junctions, that is, junctions engaged in adhesion for many hours, in which cadherin expression level is stable. After stable transfection with E-cadherin-GFP, E-cadherin dynamics were studied by 2-photon single-particle tracking and fluorescence recovery after photobleaching (FRAP) combined with 3D wide-field fluorescence microscopy, which allowed the recovery process to be analyzed from series of image stacks in the entire 3D region surrounding the photobleached volume. Image analysis of fluorescence recovery indicates that most E-cadherin did not diffuse in the membrane along mature junctions, but followed a first order turn-over process that was rate-limited by endocytosis. This movie is a z-stack (0.3 µm) of images of a FRAP experiment on MDCK cell junctions expressing E-cadherin-GFP immediately following photobleaching (CIL# 24066 shows z-series before photobleaching and CIL# 24065 shows time-lapse video of FRAP experiment). Photobleached regions are indicated by arrows. Scale bar: 10 µm.