BM neutrophils produce PGE₂by β-adrenergic stimulation. (A) Dot-plot profile for BM neutrophil sorting, May-Giemsa staining of cytospin preparation, and protein expression of mPGES-1 in BM neutrophils by western blot. Control proteins were obtained from kidneys of mPGES-1^+/+ and mPGES-1^−/− mice. Loaded protein: neutrophils 100 μg and kidneys 15 μg/lane. (B) PGE₂ levels in culture supernatants of BM neutrophils (4 h after stimulation with indicated reagents). Vehicle: PBS. Iso, isoproterenol; NE, norepinephrine; G, G-CSF. The number of each group is shown in each bar. (C) PGE₂ levels in culture supernatants of BM neutrophils (4 hours after stimulation with indicated reagents). Vehicle: PBS. β1 agonist, dobutamine; β2 agonist, clenbuterol; and β3 agonist, BRL37344 (n = 5). (D) PGE₂ levels in culture supernatants of BM neutrophils (4 hours after stimulation with indicated reagents; n = 3). Vehicle: PBS. Indo, indomethacin. (E) Deuterium-PGE₂ (PGE₂-d₁₁) levels in neutrophil culture (n = 3). Data are represented as mean ± SEM. *P < .05; **P < .01; ***P < .001.