Continued motility of self-reactive T cells on planar lipid bilayers displaying pMHC and ICAM-1. Self-reactive and influenza-specific T cells were tracked for 90 min or for as long as they were present in the imaged field. (A) Tracks of T cell paths are shown. (B) Mean velocity of individual cells was quantified. Mean motility (indicated as red bars in B) of HA-specific T cell clones was compared with self-reactive T cell clones on lipid bilayers displaying appropriate pMHC, or HA:D7 and Ob.1A12 T cells incubated on lipid bilayers displaying control CLIP-MHC. Statistical analysis was performed using a one-way analysis of variance with Tukey’s post test with * indicating P < 0.05 and ** indicating P < 0.001. For each T cell clone, 27–60 individual cells were analyzed. Each dot in the graph represents a single cell. Data are representative of at least two independent experiments.