T-cell adhesion depends on PKC-θ function. (A) Flow cytometric analysis of LFA-1 expression on wt and PKC-θ KO T cells. (B) Altered activation-dependent adhesion to ICAM-1 through LFA-1 in the PKC-θ–deficient CD4+ and CD8+ subsets. (C) Intact surface expression levels of LFA-1 in the PKC-θ–deficient CD4+ and CD8+ subsets. (D,E) Adhesion-stimulatory activities of the transiently transfected with GFP control or CA mutants PKC-θ A149E, Rap1A V12, and p110-CAAX in Jurkat (D) and primary CD3+ (E) T cells with or without PDBu/ionomycin stimulation. Results shown are the mean plus or minus SD of at least 3 independent experiments. (F,G) As experimental controls, both immunoblot for the transfected CA mutant proteins p110-CAAX and Rap1A V12, and the (p)S473 status of the PI3-K effector Akt/PKB or Rap1-GTP loading of the recombinant Rap1A V12 mutant, respectively, in lysates from unstimulated Jurkat cells are shown.