Figure 2.

Increasing TCR signal strength increases actin depletion. Confocal projections showing cell conjugates or 3-µm depth en face reconstructions across the synapse for OTI CTLs expressing Kras8⁺ probe (green) conjugated with N4 or G4 presenting EL4 for times as shown before fixation and labeling for actin (red). (a) Representative images of conjugates showing actin (red), Kras8⁺ in (green), and EL4 (blue). Maximum intensity projections (left) and en face projections (right) for merged, actin, and Kras8⁺ signals. Scale bars = 1 µm. (b) Intensity profiles for actin and Kras8⁺ along line marked between gray triangles. (c and e) Area of synapse depleted for Kras8⁺ expressed as a percentage of the total synapse area taken from the en face images for each individual cell in one representative experiment (c; n = 30 per condition) and the mean (e); bars show SEM from three separate repeats. (d and f) Area of synapse depleted for actin as a percentage of the total synapse area taken from the en face images for each individual cell in one representative experiment (d; n = 30 per condition) and the mean (f); bars show SEM from three separate repeats. A two-tailed Bonferroni-corrected Mann–Whitney test was used for statistical analysis of c–f. *, P < 0.5; **, P < 0.01.