Functional Redundancy of Class I Phosphoinositide 3-Kinase (PI3K) Isoforms in Signaling Growth Factor-Mediated Human Neutrophil Survival
Figure 6
Effect of Akt inhibition on rhGM-CSF-mediated neutrophil survival and AktSER473 phosphorylation.
Human neutrophils (5×106 cells/ml) were pre-incubated with either increasing concentrations of Akt1/2i, with 100 nm wortmannin, or with DMSO vehicle, for 20 minutes prior to treatment with 10 ng/ml rhGM-CSF or vehicle in IMDM supplemented with 10% autologous serum. A. Neutrophil were incubated with Akt1/2i at the indicated concentrations in the absence (open circles) or presence (closed circles) of rhGM-CSF 10 ng/ml. Apoptosis was assessed at 20 hours by morphology. * denotes p<0.05 for cells treated with Akt1/2i and GM-CSF versus cells treated with GM-CSF alone. B. Lysates from cells prepared as above were subjected to Western blotting for total and phosphorylated PKB. A representative immunoblot is shown. C. Densitometric analysis of n = 3 Western blots from B. *p<0.05 and **p<0.01 versus GM-CSF alone.