LAT vesicle fusion determines pLAT and pSLP76 nanoscale organization at the immunological synapse. (A–F) Jurkat cells were transfected with siCtr (A), siSyt7 (B), or siMAL (C and D) in the absence (A–C and E) or presence (D and F) of thapsigargin, and allowed to spread for 3 min on an αCD3- (A–D) or αCD45-coated (E and F) coverslip. Cells were stained for pLAT and pSLP76 and analyzed by dSTORM-TIRF imaging. Top left insets depict the correspondent zx-stack widefield image projection. Right panels show a magnified image of a region of interest (frame). pLAT in green, pSLP76 in red. Images are representative of three experiments. Bar, 10 µm.