Figure 6.

Functional assays of immune–endothelial interactions using fBMVECs. (a) Adhesion assay: fBMVECs were treated with TNF-α (20 ng/mL) for 18 h. Treatments were removed prior to addition of monocytes. Data are represented as fold difference (mean ± SEM) of adherent cells. Results show a 2.5-fold increased in adherent monocytes on activated as compared to unactivated ECs. (b) Trans-endothelial migration assay: Monocytes were added to the upper chamber of Transwell® membranes and allowed to migrate through fBMVECs towards MCP-1 in the lower chamber. Data are represented as fold difference (mean ± SEM) of migrated cells. Results show a ∼ 2-fold increase in migrated monocytes with the addition of MCP-1. (c) Representative images of the underside of the Transwell® showing migrated Calcein-AM labeled monocytes with or without MCP-1 in the lower chamber. Scare bars 100 µm.