CDC42GAP regulates both random movement and directed migration. (A) Neutrophil migration using the Boyden chamber. Migration was evaluated without stimulation, in uniform concentration, or in a gradient of 1 μM fMLP to measure chemokinesis or chemotaxis, respectively. The histogram represents the number of migrated neutrophils per field, mean ± SD; representative experiment in triplicate of 3 independent experiments is shown. (B) Neutrophil migration using transwells coated with fibrinogen. Migration was evaluated without fMLP or in uniform concentration or in a gradient of 10 μM fMLP. The histogram represents the total number of migrated neutrophils recovered from the bottom well, mean ± SD; representative experiment in triplicate from 3 independent experiments is shown (n = 9). (C) Neutrophil migration was evaluated using transendothelial migration across HUVECs in uniform concentration or in a gradient of 10 μM fMLP. The histogram represents the total number of migrated neutrophils recovered from the bottom well, mean ± SD; representative experiment in triplicate from 3 independent experiments is shown. (D) Adhesion to fibrinogen. Neutrophils were allowed to adhere to fibrinogen for 30 minutes in the absence or in the presence of 10 μM fMLP. Histogram represents the number of adherent cells counted per field, mean ± SD; representative experiment in triplicate of 3 independent experiments is shown. (E) β-2 integrin expression as assessed by flow cytometry before and after stimulation with 10 μM fMLP. The numbers indicate the relative median channel fluorescence.