Figure 5.

Glucagon inhibits CXCL-1/KC-induced actin polymerization in mice neutrophils in vitro but does not alter CD11a nor CD11b translocation to surface. Cells were pre-treated 30 min with glucagon, rolipram or vehicle in vitro and then stimulated with CXCL1/KC (10 nM) or medium for 40 min in vitro. (A) CD11a and (B) CD11b translocation to neutrophils surface was represented as the percentage of MFI values that were obtained by FACS. Representative density plots of CD11a and CD11b expression on murine BM-neutrophils surface are reported in SM Figures S3 and S4 , respectively. (C) Actin polymerization was performed by immunofluorescence with TRITC-labeled phalloidin. Representative images of actin filaments stained with TRITC-phalloidin are reported in SM Figure S5 . For (A, C), BM-neutrophils from 4 random male mice were obtained. For (B), BM-neutrophils from 5 random male mice were obtained. Each animal sample was equally distributed to all experimental groups. Sample results were obtained by blinded and randomized analysis. The vehicles used were DMSO 0.1% or medium to rolipram and glucagon, respectively. Each value represents the mean ± S.E.M. The statistical analysis was performed by one-way ANOVA followed by Newman–Keuls–Student’s t-test. Results 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.