Fig. 3From: Characterization of novel interactions with membrane NEU1 highlights new regulatory functions for the Elastin Receptor Complex in monocyte interaction with endothelial cellsBinding of EDP to the ERC potentiates monocyte adhesion to endothelial cells through NEU1. (a, b) THP-1 monocytes were incubated, or not, with κE (50 µg/mL, 1 h), labelled with Calcein-AM, and allowed to adhere for 30 min at 37 °C onto a monolayer of HUVEC pre-stimulated, or not, with PMA (100 nM). Cells were then washed, fixed and visualized under a fluorescent microscope. a A representative field is shown. b The percentage of surface area covered by adherent monocytes was calculated for each condition from 10 different fields per experiment, each run in duplicate. Results were expressed as mean ± SEM of 4–8 independent experiments and normalized to the control condition (without, w/o) in presence of PMA. Statistical analysis was performed by Student’s t-test (***p < 0.001; ns: non-significant). c Similar experiments as in b except that HUVEC, instead of THP-1 cells, were incubated, or not, with κE (50 µg/mL, 1 h) prior to monocyte adhesion. Results were expressed as mean ± SEM of 3–4 independent experiments. Statistical analysis was performed by Student’s t-test (***p < 0.001; ns: non-significant). d THP-1 monocytes were incubated, or not, with DANA (400 µM), anti-CD18 blocking antibody (10 µg/mL) or isotype control (10 µg/mL) for 1 h at 37 °C before stimulation, or not, by κE (50 µg/mL), V14 + κE (molar ratio 2:1) or V14 peptide alone for 1 h at 37 °C. After adhesion (30 min, 37 °C), the percentage of surface area covered by adherent monocytes was calculated for each condition from 10 different fields per experiment, each run in duplicate. Results were expressed as mean ± SEM of 3–4 independent experiments, and normalized to the control condition (w/o). Statistical analysis was performed by Student’s t-test (**p < 0.01; ***p < 0.001; ns: non-significant). Black bars, non-stimulated cells; white bars, κE-stimulated cells. e Similar experiments as in d except that HUVEC, instead of THP-1 cells, were incubated, or not, with DANA (400 µM) followed by stimulation, or not, κE (50 µg/mL), V14 + κE (molar ratio 2:1) or V14 peptide alone for 1 h at 37 °C. Results were expressed as mean ± SEM of 3–6 independent experiments. Statistical analysis was performed by Student’s t-test (***p < 0.001; ns: non-significant)Back to article page