Fig. 4

CD11b and endothelial adhesion modulation by semaglutide. A Work flow of the assay based on neutrophil-like dHL-60 cells treatment with semaglutide (Sema) 1 nM for 2 h and following analysis a) migration on a Sun-Chip or b) treatment with fMLP (10 μM, 90 min) and CD11b analysis by flow cytometry. Bar graph with individual data points represents the CD11b levels after treatments [n = 6; one-way ANOVA, F(2,5) = 9.817, p = .0236; p < 0.05; Dunett’s post-hoc fMLP (270 ± 97.04) vs Sema (190.5 ± 46.72), adjusted p value = 0.0433; p < 0.05]. Representative histogram of flow cytometry analysis or Sun-Chip migration assay. B Work flow of the assay based on neutrophil-like dHL-60 cells with fMLP (10 μM), with or without Sema treatment at 1 nM, 100 nM or 1000 nM, treatment for 90 min and its adhesion to aortic endothelial cells (HAEC) monolayer after high glucose (HG) treatment for 30 min. Representative fluorescence microscopy images are shown (HAEC in red, dHL-60 cells, in green. The nuclei are counterstained with DAPI. Scale bar = 100 μm). Bar graph with individual data points shows the analysis based on ChemiDoc [n = 5, Veh + fMLP (100 ± 0) vs Sema (1 nM) + fMLP (84.62 ± 8.37); t(4) = 4.106, p = .0148; p < 0.05] or microscope images [n = 5, Veh + fMLP (100 ± 0) vs Sema (100 nM) + fMLP (0.769 ± 0.185); t(4) = 2.780, p = 0.0498; p < 0.05]