Effects of AG490 and PD98059 on phosphorylation of STAT3 and ERK in neutrophils stimulated by G-CSF.
(A) Differential expression of STAT3 isoforms in neutrophils and mononuclear cells (MNC). The cell lysates from an equal number of cells (2.5 × 106) were analyzed by immunoblotting using antibody against STAT3. The results shown are representative of 3 independent experiments. (B) Neutrophils were preincubated with AG490 (50 μM) for 20 minutes, and thereafter stimulated with G-CSF (50 ng/mL) for 10 minutes at 37°C. The cell lysates from an equal number of cells (2.8 × 106) were analyzed by immunoblotting using antibodies against phosphorylated forms of STAT3 and ERK. The equal loading of proteins onto each lane was confirmed by immunoblotting using antibody against p38. The results shown are representative of 3 independent experiments.
