CD2–CD58 interaction is organized in membrane microdomains that are generated and transported by actin filaments. (A) A schematic of a Jurkat T cell spreading on CD58 bilayer forming CD2–CD58-enriched microdomains in the actin-rich lamellae, corresponding to the boxed region in B. (B) Spinning-disk confocal fluorescence microscopy of Jurkat cells expressing actin-GFP (red) that spread on CD58 (green)-containing bilayer imaged at the cell–bilayer interface. (C) Dynamics of CD58 microdomains. The t = 0 s time was defined as the time at which the T cell had just adhered to the bilayer and some accumulation of CD58 under the cell was observed. But before the cell began to spread (t = ∼45 s), microdomains were formed at the initial cell–bilayer contact under cortical actin filaments. After the cell spreads, new microdomains were continuously formed at cell periphery where actin filaments are polymerizing (t = ∼130 s). Microdomains were transported to the cell center and formed a larger central cluster (cSMAC-like structure; t = ∼340 s). (D) Trajectories of individual CD2–CD58 microdomains transported from the cell periphery to the cell center are shown. (E) A histogram of the speeds of CD2–CD58 microdomain transport is shown. Bars, 5 µm.