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. 2005 Oct;167(4):1173–1181. doi: 10.1016/S0002-9440(10)61205-9

Figure 6.

Figure 6

Inhibition of FH and FH15-20 binding to HUVECs by heparin or mAb. To analyze the role of heparin-binding sites in binding of FH and FH15-20 to HUVECs we preincubated FH (A) or FH15-20 (B) with heparin (3, 30, and 300 μg/ml), or FH15-20 with dextran sulfate (DS; 3, 30, and 300 μg/ml) (C) before adding the mixture to the cells and the subsequent flow cytometric analysis. In each test a cell preparate treated without FH or FH15-20 was used as a negative control and is indicated as control. D: To analyze the effect of the carboxy-terminally binding anti-FH mAb 3D11, FH15-20 was preincubated with the Fab fragment before adding the mixture onto HUVECs and subsequent flow cytometric analysis. E and F: To further study the effects of 3D11 and C3d on the binding of FH15-20 to HUVECs, a range of concentration of 3D11 and C3d was used and the percentage of specific mean fluorescence intensity (MFI) is shown. The bound proteins were detected using monoclonal mouse anti-FH antibody (IXF9) and a fluorescein isothiocyanate-labeled secondary antibody. For the experiment in the presence of Fab fragment of 3D11 and C3d we used polyclonal anti-FH antibody instead of IXF9.