Volume 119, Issue 6, Pages 1708-1719 (December 2000) Sterol carrier protein 2 gene transfer changes lipid metabolism and enterohepatic sterol circulation in mice  Silvana Zanlungo, *, Ludwig Amigo, *, Hegaly Mendoza, *, Juan Francisco Miquel, *, Carlos Vío, ‡, Jane M. Glick, §, Annabelle Rodríguez, ∥, Karen Kozarsky, ¶, Verónica Quiñones, *, Attilio Rigotti, *, Flavio Nervi, *  Gastroenterology  Volume 119, Issue 6, Pages 1708-1719 (December 2000) DOI: 10.1053/gast.2000.20198 Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 1 Immunoblot analysis of SCP-2 expression in livers of mice infected with SCP-2 recombinant adenovirus. Liver homogenates were prepared and subjected to 12% SDS-PAGE and Western blotting with anti–SCP-2 and antialbumin antibodies. The change in SCP-2 protein expression of Ad.rSCP-2–infected mice compared with control AdE1Δ–infected mice is shown after correction for albumin signal. Gastroenterology 2000 119, 1708-1719DOI: (10.1053/gast.2000.20198) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 2 Immunohistochemical localization of SCP-2 in liver tissue of SCP-2 recombinant adenovirus–infected mice. Low-resolution immunohistochemistry was performed in liver sections of (A and C) Ad.rSCP2- and (B and D) AdE1Δ-infected mice. (E and F) High-resolution hepatic immunohistochemistry of Ad.rSCP2-infected mice. Bar = 50 μm (A and B); bar = 25 μm (C and D); bar = 10 μm (E); bar = 5 μm (F). Gastroenterology 2000 119, 1708-1719DOI: (10.1053/gast.2000.20198) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 3 Plasma lipoprotein cholesterol distribution in mice with adenovirus-mediated SCP-2 gene transfer. Plasma lipoproteins were separated by fast protein liquid chromatography gel filtration, and cholesterol was measured in lipoprotein fractions. Values are expressed as percentages of total plasma cholesterol for 5–6 nonfasted mice in each experimental group. *P < 0.001. Gastroenterology 2000 119, 1708-1719DOI: (10.1053/gast.2000.20198) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 4 Hepatic expression of apoA-I, apoB, and apoE in Ad.rSCP2-infected mice. Total hepatic RNA was prepared, electrophoresed, and transferred to nylon membranes. Apolipoprotein gene expression was evaluated by RNA blot hybridization with 32P-labeled cDNA probes and densitometric analysis. The fold change in mRNA expression in Ad.rSCP-2 mice compared with AdE1Δ-infected mice is shown after normalization against GAPDH mRNA. AP < 0.05. Gastroenterology 2000 119, 1708-1719DOI: (10.1053/gast.2000.20198) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 5 Hepatic LDL receptor and SR-BI expression in SCP-2–overexpressing mice. Total membrane extracts were prepared, size fractionated by SDS-PAGE, immunoblotted with anti–SR-BI and LDL receptor antibodies, and subjected to densitometric analysis. Anti-ϵCOP antibody was used for membrane protein loading control. The fold change in receptor expression of Ad.rSCP-2 mice compared with control Ad.E1Δ mice is shown after correction for ϵCOP signal. AP < 0.05. Gastroenterology 2000 119, 1708-1719DOI: (10.1053/gast.2000.20198) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 6 Hepatic expression of cholesterol 7α-hydroxylase and oxysterol 7α-hydroxylase in Ad.rSCP2-infected mice. Hepatic RNA was prepared, electrophoresed, and transferred to nylon membranes. Hydroxylase gene expression was evaluated by RNA blot hybridization with 32P-labeled cDNA probes and densitometric analysis. The fold change in mRNA expression of Ad.rSCP-2 mice compared with AdE1Δ-infected mice is shown after normalization against GAPDH mRNA. AP < 0.05. Gastroenterology 2000 119, 1708-1719DOI: (10.1053/gast.2000.20198) Copyright © 2000 American Gastroenterological Association Terms and Conditions