Video 1. Example of a WT control T cell undergoing transendothelial migration imaged by time-lapse confocal microscopy.
Download video file (452.2KB, mp4)
Control and FMNL1 KO T cells were activated ex-vivo and differentially dye-labeled with CFSE or CellTrace Yellow (CTY). These T cells were mixed at a 1:1 ratio and perfused into flow chambers containing bEnd.3 brain endothelial cell monolayers and kept under shear flow (2 dyne/cm2) for up to 30 min. During this time, phase contrast and fluorescence images were acquired every 20 s using a spinning-disk confocal microscope. Phase contrast (left), CTY fluorescence (middle), and overlay (right) images are shown. This representative control T cell completes transendothelial migration as shown by the progressive disappearance of the white phase contrast ring around the T cell. Time is displayed as min:s.