FIGURE 7.

ApoA-I nitrated on tyrosine 166 obtained from the normal aortic tissue is not on an HDL-like particle and behaves similarly to that isolated from atherosclerotic lesions. Aortic tissue that was histopathologically determined to appear normal was used to prepare a tissue homogenate as described under “Experimental Procedures.” A, aortic tissue homogenates from normal and atherosclerotic human artery wall tissue (n = 5 each) with the indicated amounts of protein were fractionated by gradient (5–15%) SDS-PAGE and stained with SYPRO Ruby Red. B, Western blot of duplicate gel as in A with the indicated amount of protein (15 μg) per lane (n = 5) probed with anti-NO₂-Tyr¹⁶⁶-apoA-I mAb 4G11.2. C, recovery of apoA-I (μg) per gram of tissue (wet weight) from the normal aortic tissue and from atherosclerotic lesion tissue was determined from quantitative Western blot analysis of apoA-I-immunoreactive bands and total apoA-I content as described under “Experimental Procedures” and is qualitatively presented in Figs. 8B and 4B. D, percentage of NO₂-Tyr¹⁶⁶-apoA-I to total apoA-I present in the starting material as indicated was quantified as in C. Values were determined from n = 5 samples; error bars represent ±S.D.