Figure 1.

BMP9 and BMP10 increase monocyte recruitment to TNFα-stimulated HAECs in a concentration-dependent manner. HAECs were treated with BMP9 and BMP10 16 h prior to TNFα treatment (0.05 ng/ml, 4 h). Monocytes were perfused over TNFα-stimulated HAECs in a flow adhesion assay in the presence of media alone, BMP9 or BMP10. A, representative images of HAEC monolayers that were untreated, or treated with BMP9 (5 ng/ml), BMP10 (5 ng/ml), TNFα, BMP9 + TNFα, or BMP10 + TNFα. Adherent monocytes are the bright phase cells (white arrow) and transmigrated monocytes shown as the smaller dark phase cells (black arrow). Experiments were performed in triplicate and the data are representative of n = 3 biological repeats. B and C, concentration-response analysis of the recruitment of monocytes to HAEC monolayers, in the presence or absence of TNFα, with increasing concentrations of BMP9 (B) (0–5 ng/ml) and BMP10 (C) (0–5 ng/ml). D and E, monocyte behavior (rolling, clear bar; adherence, gray bar; and transmigration, black bar) was expressed as a percentage of total recruitment to TNFα-stimulated HAECs in the presence of BMP9 (D) and BMP10 (E). F, analysis of the recruitment of monocytes to BOEC monolayers, treated with 5 ng/ml of BMP9 or BMP10, in the presence or absence of TNFα. G, monocyte behavior (rolling, clear bar; adherence, gray bar; and transmigration, black bar) was expressed as a percentage of total recruitment to TNFα-stimulated BOECs in the presence of BMP9 or BMP10. Error bars represent ± S.E. *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001.