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Arterioscler Thromb Vasc Biol

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1 Arterioscler Thromb Vasc Biol
Effects of Genotype and Diet on Cholesterol Efflux into Plasma and Lipoproteins of Normal, Apolipoprotein A-I-, and Apolipoprotein E-Deficient Mice by Yadong Huang, Yanhong Zhu, Claus Langer, Martin Raabe, Shili Wu, Bernd Wiesenhütter, Udo Seedorf, Nobuyo Maeda, Gerd Assmann, and Arnold von Eckardstein Arterioscler Thromb Vasc Biol Volume 17(10): October 1, 1997 Copyright © American Heart Association, Inc. All rights reserved.

2 Electrophoretic demonstration of lipoproteins in plasmas of normal, apoA-I- and apoE-deficient mice.
Electrophoretic demonstration of lipoproteins in plasmas of normal, apoA-I- and apoE-deficient mice. Aliquots of 1 μL of mouse plasma were electrophoresed in a 1% agarose gel. Lipoproteins were then demonstrated by staining with Fat Red 7B (A), by anti-apoA-I immunoblotting (B), or anti-apoE immunoblotting (C). Lanes 1 and 2 contain normal mouse plasma, lanes 3 and 4 contain apoA-I-deficient plasma, lanes 5 and 6 contain apoE-deficient plasma. Lanes 1, 3, and 5 present lipoproteins in plasmas of CD-fed animals and lanes 2, 4, and 6 lipoproteins in plasmas of FD-fed animals. Yadong Huang et al. Arterioscler Thromb Vasc Biol. 1997;17: Copyright © American Heart Association, Inc. All rights reserved.

3 2D-PAGGE and immunoblotting of apoE-containing lipoproteins in different mouse plasmas.
2D-PAGGE and immunoblotting of apoE-containing lipoproteins in different mouse plasmas. Nondenaturing 2D-PAGGE was performed in the sequence agarose gel electrophoresis→nondena-turing polyacrylamide gel electrophoresis. After electroblotting to nitrocellulose membranes, apoE-containing lipoproteins were detected with biotinylated polyclonal goat antibodies against human apoE and streptavidin-biotinylated horseradish peroxidase complex. Parts a and b present electrophoretograms of normal mouse plasma, parts c and d electrophoretograms of apoA-I-deficient plasma, parts e and f electrophoretograms of apoE-deficient plasma. Plasmas of CD-fed animals have been separated for parts a, c, and e and plasmas of FD-fed animals for parts b, d, and f. ApoE-containing lipoproteins of normal and apoA-I-deficient mice with pre-β mobility in agarose gels (cf. Fig. 1, lanes 2 and 4) are not detectable in the polyacrylamide gel. They probably correspond to VLDL, which because of their size do not migrate in polyacrylamide gels. Yadong Huang et al. Arterioscler Thromb Vasc Biol. 1997;17: Copyright © American Heart Association, Inc. All rights reserved.

4 Dietary effects on the uptake of cell-derived 3H-cholesterol by various lipoproteins in plasmas of normal, apoA-I-deficient, and apoE-deficient mice. Dietary effects on the uptake of cell-derived 3H-cholesterol by various lipoproteins in plasmas of normal, apoA-I-deficient, and apoE-deficient mice. Mouse plasmas were incubated for 1 minute with 3H-cholesterol-labeled normal human fibroblasts. Aliquots of 20 μL of plasma were then electrophoresed in 0.75% agarose gels. After electrophoresis, each lane of the gel was cut into 5-mm-long segments. The lipids in each slice were extracted with chloroform/methanol (2:1), and their radioactivity was determined by liquid scintillation spectrometry. The values present means and standard deviations of three experiments. Yadong Huang et al. Arterioscler Thromb Vasc Biol. 1997;17: Copyright © American Heart Association, Inc. All rights reserved.


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