Fig. 6: CCR2 ligands are secreted but do not bind to HUVECs. | Nature Communications

Fig. 6: CCR2 ligands are secreted but do not bind to HUVECs.

From: Migration arrest and transendothelial trafficking of human pathogenic-like Th17 cells are mediated by differentially positioned chemokines

Fig. 6

a Culture medium collected from the upper and lower wells of transwells with HUVEC monolayers either not stimulated (NS) or stimulated with TNF-α + IFN-γ were assayed for CCL2, CCL7, and CCL8 by ELISA (n = 3 separate experiments). Each symbol shows data from one experiment, with assays done in duplicate, and bars indicate means ± SEM. b Confocal microscopy images at × 40 magnification of permeabilized and non-permeabilized TNF-α + IFN-γ-stimulated HUVECs immunostained for CCL2, CCL7, CCL8, CCL5, CCL20, and CXCL9 (green) and stained using phalloidin for polymerized actin (magenta) and DAPI for nuclei (blue). The scale bars indicate 10 µm. Images are representative of three experiments. c Biotinylated CCL5 (0.1 μM) and CCL2 (1 μM) were incubated with CHO-K1 cells and the heparan-sulfate deficient cell line, D-677. Chemokine binding was detected using streptavidin-phycoerythrin (PE) and flow cytometry. Data shown are representative of three separate experiments.

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