CCL5-mediated PLD activation regulates T-cell migration. (A) Activated PB T cells were pretreated with either ethanol (carrier) or the specified inhibitors for 1 hour at the concentrations indicated, and CCL5-mediated chemotaxis was measured using 10 nM CCL5. The data are presented as percentage migration, with the number of migrated cells at 10 nM CCL5 taken as 100%. Data representative of 3 independent experiments are shown (± SD); *P < .05. (B) T cells were pretreated with either ethanol (carrier), 500 μM 2,3-DPG, or 0.1% 1-butanol for 1 hour prior to 15-minute treatment with 10 nM CCL5. Cells were harvested and lysates resolved by SDS-PAGE and immunoblotted with anti–phospho-4E-BP1 (Thr 37/46) antibody. Blots were stripped and reprobed with anti–4E-BP1 antibody as a loading control. The relative fold increase of 4E-BP1 phosphorylation is shown as signal intensity over loading control. Data are representative of 2 independent experiments.