Western blot analysis of vasculin and SSH6-β. For each sample, amounts equivalent to 25 μg protein were applied. Vasculin and SSH6-β were detected by scFv-2A4 (5 μg/mL) and visualized using enhanced chemiluminescence. (A) Analysis of a variety of human cell lines and tissues. Lane 1 shows atherosclerotic plaque; lane 2, human plasma; lanes 3 to 6, human cell lines (3, colon carcinoma; 4, human umbilical vein endothelial cells; 5, Louis lung carcinoma; 6, vascular SMC line); lanes 7 to 11, human tissues (7, muscle; 8, brain; 9, ovary; 10, aorta); and M, marker. (B) Plasma samples from patients with (lane 1 to 5) or without (lane 6 to 10) clinical manifestations of renal atherosclerotic disease. M indicates marker. (C) A 2D gel electrophoresis analysis of a human atherosclerotic plaque lysate. Advanced plaque lysate (50 μg) was separated using a 7-cm pH gradient strip (pH 4-7) and a 12% SDS-PAGE and transferred onto nitrocellulose.