A) Concentration-dependent effect of fMLP on migration of dHL60 cells through a transmembrane filter. An increased concentration of fMLP was added to the bottom wells while the dHL60 cells were added on to the top plates. dHL60 cell migration was quantitated by MPO activity of the bottom wells. Results are presented as the mean ± SEM from 4 separate experiments. ** p < 0.01 vs. buffer control by Fisher's LSD test. B) Effect of TAT-Pyk2-CT on chemotaxis of dHL-60 cells to 10-8 M fMLP. dHL-60 cells were preincubated with TAT-Pyk2-CT or TAT-GFP and were added on the top plate of the migration system. dHL-60 cell migration in response to 10-8 M fMLP was measured after 90 min. Results are presented as the mean ± SEM from 4 separate experiments. *P < 0.05 and **p < 0.01 vs. fMLP alone by Fisher's LSD test. C) Effect of TAT-Pyk2-CT on autophosphorylation of Pyk2 induced by fMLP in adherent dHL60 cells. dHL60 cells were preincubated with 2 μM TAT-Pyk2-CT or 2 μM TAT-GFP for 30 min, and then added to Fg-coated plates with 1 μM fMLP for 10 min. Pyk2 phosphorylation was detected by Y402 phosphorylation-specific antibody (upper panel). Total Pyk2 expression was detected by anti-Pyk2 antibody (lower panel). Representative blot is shown (n = 3).