FIGURE 3.

LZTFL1 transiently redistributes to the contact zone between T cells and APCs. (A) Jurkat T cells (denoted by T) were copelleted with BMQC-stained, SEE-loaded Raji B cells (blue; denoted by B) and incubated at 37°C for 1.5-, 5-, and 15-min increments to induce IS formation. Cell conjugates were fixed and co-stained with Abs for LZTFL1 (green) and TCRβ (red). SEE-, no SEE-loading control, incubated for 15 min. Arrows indicate the T cell and B cell contact zone. (B) The percentage of conjugated T cells was calculated for all T cells in 10 fields chosen at random at indicated time points (mean ± SD). Results are representative of three independent experiments. *, p-value < 0.05. (C) LZTFL1 accumulation at the IS and the distal pole (DP) was quantified, and the percentage in total conjugates was plotted for both (mean ± SD). Results are representative of three independent experiments. *, p-value < 0.05 (comparison to LZTFL1 located at the IS after 1.5 min of incubation); (D and E) Live cell imaging of LZTFL1 localization at the IS. Jurkat T cells (red; denoted by T) stably expressing Halo-STOP (D) or Halo-LZTFL1 (E) interacting with SEE superantigen–loaded Raji B cells (blue; denoted by B) and forming the IS at indicated times on live cell imaging, as analyzed by wide field microscope (Supplemental video1). Data are representative of two independent experiments. (Scale bar, 10 μm.)