DM enhancement of T-cell migration is a GCR- and Lck-dependent event. (A) Primary human T cells were labeled with calcein-AM and incubated in the presence or absence of 5 μM mifepristone (RU486) for 1 hour. The cells were then treated with DM (1 μM) for 2 hours after which the cells were examined for their ability to migrate in response to CXCL12 (100 ng/mL) for 2 hours. (B) Primary T cells were treated with damnacanthal (500 nM for 1 hour), a Lck inhibitor, genistein (50 μM for 1 hour), a tyrosine kinase inhibitor, lavendustin-A (10 μM for 10 minutes), a Src kinase inhibitor, staurosporine (200 nM for 30 minutes), a protein kinase C inhibitor, or wortmannin (20 nM for 30 minutes), a phosphatidylinositol 3 kinase inhibitor. The dose and timing of each inhibitor treatment were previously optimized. The cells were then treated with 1 μM DM for 2 hours followed by a migration assay. The vehicle for all the treatment was maintained in parallel. No significant alterations in cell viability were detected for any of these inhibitors or vehicle as assessed by Trypan blue exclusion (data not shown). In all panels, the fluorescent signal of migrated cell was measured using a microfluorimeter. Triplicate well determinations were performed for each treatment group. The data were presented represent the average of 3 individual experiments plus or minus SE. *Significant value (P < .05). PMI indicates the percent migration index.