Figure 3

Polarization of γ-tubulin towards IS in CD4+ T cells upon engagement of TCR. (A) Sample images show absence (−) and presence (+) of γ-tubulin oriented towards IS in Id-specific T cells conjugated to Id positive F9 cells, displaying polarized F-actin and Id-specific TCR. (B) Histogram shows γ-tubulin polarization ratio in Id-specific T:APC conjugates displaying F-actin and Id-specific TCR polarization to the synapse, using T cells from either Sh2d2a+/+ or Sh2d2a−/− mice, after 30 minutes incubation. The stippled line indicates the threshold value of 1.5. Conjugates with polarization ratio >1.5 were defined as having γ-tubulin polarized to IS. Data are from one experiment. Statistical significance was assessed using Mann-Whitney test. (C) Graph shows frequency of Id-specific T:APC conjugates displaying γ-tubulin polarization to the synapse. (D) Graph shows median γ-tubulin polarization ratios towards synapse in the presence of Id positive F9 cells. Data in C and D represent median values (+95% CI) of three independent experiments, n > 50 gated events in each experiment. Statistical significance was assessed using paired student’s t-test.