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. Author manuscript; available in PMC: 2012 Aug 13.
Published in final edited form as: Curr Biol. 2003 Sep 16;13(18):1619–1624. doi: 10.1016/j.cub.2003.08.005

Figure 4. Itk Is Required for Vav Recruitment and Cdc42 Activation at the Immune Synapse.

Figure 4

(A) Conjugates were labeled with recombinant GFP-WASP-GBD to detect Cdc42-GTP. Note the enrichment of activated Cdc42 at the immune synapse of the conjugate formed with the wt T cell, but not the Rlk−/− × Itk−/− T cell.

(B) Randomly chosen conjugates formed in the absence (hatched bar) or presence (solid bars) of Ag were scored for accumulation of Cdc42-GTP at the immune synapse.

(C) Double labeling was performed by using the GFP-WASP-GBD reagent to detect Cdc42-GTP (green) and mAb 26E6 to detect the open, active conformation of WASP (red). The overlaid image shows extensive colocalization of activated Cdc42 and WASP at the immune synapse (yellow).

(D) Conjugates were labeled with anti-Vav antibody. Note the enrichment of Vav at the immune synapse of the conjugate formed with the wt T cell, but not the Rlk−/− × Itk−/− T cell.

(E) Randomly chosen conjugates formed in the absence (hatched bar) or presence (solid bars) of Ag were scored for accumulation of Vav at the immune synapse. Data in (B) and (F) represent means from at least three independent experiments ± SD (the asterisk indicates a significant difference from wt + Ag, p < 0.001).