Figure 6.
Effects of LRP1 (low-density lipoprotein-related protein 1) and detection of versikine in human ascending aortic aneurysms. A, LRP1 localization in the ascending aorta of mice lacking the catalytic domain of a disintegrin and metalloproteinase with thrombospondin motifs 5 (Adamts5Δcat mice). Colocalization of LRP1 (Alexa 647, displayed in red) and αSMA (alpha smooth muscle actin; Alexa 568, displayed in green) in the mouse aorta as visualized by immunofluorescence. Elastin fibers are in white (autofluorescence captured with 488 nm laser excitation). Control sections stained with isotope IgGs. Magnification ×60, scale bars=10 μm. B, Relative gene expression of ADAMTS1, LRP1, ADAMTS5, and VCAN in human aortic smooth muscle cells (SMCs) treated for 72 h with LRP1 siRNA (n=3). β-actin was used as reference gene. Data points are mean±SD. Statistical significance was calculated using paired Student t tests. *P<0.05. C, Immunoblotting performed on cell lysate and conditioned media after transfection (n=3). Silencing LRP1 results in a decrease in versikine and an increase in ADAMTS-1 in the conditioned media. D, Localization of versican (Alexa568, displayed in green) and versikine (Alexa 647, displayed in red) in human ascending aortic aneurysms as visualized by immunofluorescence. Magnification ×20, scale bar=200 μm and ×60, scale bar=20 μm.