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. 2022 May 20;23(10):5705. doi: 10.3390/ijms23105705

Figure 5.

Figure 5

CD47-mediated signals in vascular endothelial cells inhibit IL-17-induced VCAM-1 expression and leukocyte adhesion. Primary cultures of vascular endothelial cells from WT mice were treated overnight with IL-17 (10 ng/mL) alone or in the presence of CD47-binding 4N1K or control 4NGG peptides (10 nM). (a) To determine VCAM-1 expression, cells immunostained for VCAM-1 (red) (200×) were imaged and analyzed using ImageJ software, as described in Materials and Methods. Nuclei were stained with DAPI (blue). Results are shown as the mean fluorescence staining relative to untreated cells ± SEM; (b) To determine leukocyte adhesion, treated cells were washed gently and overlaid with CFSE-labeled leukocytes, as described in Materials and Methods, and after washing away non-adherent cells, fluorescence intensity of adherent cells was measured using a fluorescence microplate reader. Data are presented as the mean fluorescence intensity of adherent cells relative to untreated cells ± SEM. n = 3–4 each treatment. * p < 0.05.