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Data Supplement for Arteriosclerosis, Thrombosis, and Vascular Biology: Volume 21, Issue 1 -- Page 88
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Data Files
Figure I (JPEG) A, Northern blot analysis of human apo(a) expression. Total RNA was extracted from the liver, aorta, spleen, and bone marrow, which are enriched in blood cells. Northern blotting was performed using a human apo(a) cDNA probe. Rehybridization of the membrane with a human β-actin probe showed that relatively equal amounts of RNA had been loaded in each lane. B and C, Immunoblotting analysis of transgenic rabbit plasma apo(a). Aliquots of plasma were separated by either 4% nondenaturing polyacrylamide gel electrophoresis (B) or 4% SDS-PAGE under nonreducing or reducing conditions (C). Almost all of the plasma apo(a) in cholesterol-fed transgenic rabbits was associated with apoB. The plasma of chow-fed homozygous WHHL transgenic rabbits at age of 4 months with plasma cholesterol ~700 mg/dL and human apo(a) ~42 nM are used to show both free apo(a) and Lp(a) complex. After electrophoretic transfer, the proteins were immunoblotted using anti-human apo(a) mAb. The same immunoblot membranes were stripped and reprobed with anti-apoB polyclonal Ab. *high molecular weight Lp(a)-like particles, presumably bound by covalent bonds; **low molecular weight Lp(a)-like particles bound by noncovalent bonds. Arrows indicate 2 kinds of isoforms of apo(a) molecules. Prestained SDS-PAGE marker (Bio-Rad, Hercules, Cal) was loaded in SDS-PAGE under the reducing condition to indicate the molecular weight.
Figure II (JPEG) Quantitation of cholesterol content in plasma lipoproteins. Density gradient fractions from chow-fed (upper panel) and cholesterol-fed (lower panel) male rabbits were collected by ultracentrifugation and cholesterol contents were quantitated as described. The combined recovery for each animal averaged ~80% of the total amount in plasma.
Prepared by: the Data Supplement Manager
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