CIL:24069

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 z-stack (0.3 µm) of images is of a FRAP experiment on MCF7 cell junctions expressing E-cadherin-GFP before photobleaching. Also available is the z-stack immediately following photobleaching (CIL# 24070) and the time-lapse video of FRAP experiment (CIL# 24068). Photobleached regions are indicated by arrows. Scale bar: 10 µm.