Figure 5

MCP-1 in the plasma and heart/aortic tissues in human apoB100 transgenic mice on high fat diet and exposed to SSW: (A) Levels of MCP-1 in the plasma of smoking mice at two different time points during smoking. 30 μl of cytokine standard or plasma sample were added into the appropriate wells of a Mouse Cytokine array (Quansys Biosciences, Utah, USA) containing an antibody to this chemokine. The images were captured by a CCD camera and analyzed using the Quansys image analysis software. The concentration of MCP-1 was calculated and plotted using a standard curve. (B) Immunoblot analysis for MCP-1 in the heart/aortic tissue (the upper half of the heart together with ascending aorta). The protein extractions were quantified using a DC protein assay kit (Bio-Rad, Hercules, CA) and equal amounts of protein were loaded in each lane of SDS-Polyacrylamide Gel. A 45 KD protein was used to evaluate equal loadings because this protein is always constant, independent of the treatment when the gels are stained with Commasie blue. (C): Immunolabeling for MCP-1 in an arterial vessel of the heart tissue shows that this chemokine is produced by the endothelial cells (arrowheads) and by some of the cells in the adventitia (arrow). The mice were exposed to SSW periodically for 1 year. Representative images of one mouse out of three.