Redistribution of Akt2 but not Akt1 in polarized neutrophils. Bone marrow–derived neutrophils from WT, Akt1^−/−, and Akt2^−/− mice were seeded onto glass coverslips and incubated with buffer (A) or 1μM fMLF (B) for 2 minutes at 37°C. Stimulation was terminated when 1 mL of ice-cold PBS was added. The cells were fixed with paraformaldehyde and stained first with a pan-Akt mouse mAb and then with an Alexa Fluor 488–conjugated goat anti–mouse IgG together with rhodamine phalloidin (1:500, to stain F-actin). 4′,6-Diamidino-2-phenylindole (in mounting solution) was used for nuclear staining. Representative images shown are selected from 20 images for each experimental condition from 2 independent experiments.