CIL:24065

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 is a time-lapse video of a FRAP experiment on MDCK cell junctions expressing E-cadherin-GFP (each image is a projection of 14 z-frames). Also available are z-stacks (0.3 µm) of images of the FRAP experiment before (CIL# 24066) and immediately following photobleaching (CIL# 24067). Photobleached regions are indicated by arrows. Scale bar: 10 µm.