Figure 4.

Glucagon inhibits mice neutrophils chemotaxis and ROS production in vitro. (A) Representative sample of GcgR and β-actin expression in neutrophils obtained from mice BM. The expressions of GcgR and β-actin were determined by western blot. Results are representative of 3 animals. Full-length blots of GcgR and β-actin are reported in SM Figure S2 . (B–D) Effect of glucagon on CXCL1/KC- (25nM), PAF- (20 µM), or fMLP-induced (1 µM) chemotaxis of neutrophils isolated from mice BM in vitro, respectively. Cells were pretreated 30 min with glucagon, rolipram or vehicle in vitro and then let to migrate for 40 min towards stimulus in a chemotaxis chamber. (E) ROS production by BM-neutrophils in vitro. Cells were pretreated 30 min with glucagon, rolipram or vehicle in vitro and then stimulated with zymosan A (0.1 mg/mL) for 1 h. The vehicles used were DMSO 0.1% or medium to rolipram and glucagon, respectively. Each value represents the mean ± S.E.M. from 5 animals. The statistical analysis was performed by one-way ANOVA, followed by Newman–Keuls–Student’s t-test. Results from (B–E) are representative of three individual assays. Results from (A) are representative of one individual assay. ⁺ P < 0.05 compared to non-stimulated cells. ^* P < 0.05 compared to stimulated cells. Gcg, Glucagon; Rol, Rolipram.