FIG. 4.

PLD activity is required for TCR-stimulated lymphocyte adhesion to ICAM-1. (A) Jurkat T cells transfected with the indicated Rap1 construct tagged with GFP were plated on ICAM-1 and stimulated with or without anti-CD3 antibodies, and adherent, transduced cells were measured with a fluorescent plate reader. Untransfected Jurkat cells (B) or CD4⁺ splenocytes from C57BL/6 mice (C) were labeled with 5-carboxyfluorescein, treated with or without t-butanol or n-butanol, and stimulated with or without anti-CD3 antibodies, and adhesion to ICAM-1 was measured as in panel A. (D) Jurkat cells labeled with 5-carboxyfluorescein and transfected with the indicated shRNA were stimulated with or without anti-CD3 antibodies, and adhesion to ICAM-1 was measured as in panel A. Results are shown as means ± SEM (n = 4), except in panel C, where the results of an experiment representative of two are shown.