Fig. 3.

HNP1 interacts with lipoproteins enriched in VLDL and LDL with high affinity. (A) Apoe^−/− mice were fed a high fat diet and injected with HNP1 (10 μg, i.v.) and plasma was collected after 24 h. FPLC-assisted fractionation of plasma lipids is displayed (top). Lipid fractions of HNP1-treated mice were spotted on a nitrocellulose membrane and probed with antibodies to HNP1, ApoA1, ApoB, and ApoC3 (bottom). (B/C) Surface plasmon resonance reveals interaction between HNP1 and human apolipoproteins ApoC3 or ApoB (B) or human plasma lipoproteins (hLDL or hHDL) (C). Indicated apolipoproteins or plasma lipoproteins were perfused over a biacore sensor chip coated with HNP1 and the resulting response was assessed. (D–F) Precipitation of lipid-bound HNP1 reduces plasma cholesterol levels. Experimental outline is detailed in (D). Please note the definition of samples (i), (ii), (iii), (iv). Cholesterol levels obtained at different steps (i), (ii), (iii), and (iv) of precipitation (E). Precipitated samples were spotted onto a nitrocellulose membrane and probed with indicated antibodies (F).