FVIII clearance after adenovirus-mediated overexpression of human LDLR in mice. (A) Liver membrane extracts of wild-type mice that received Ad-CMV-β-Gal (n = 2) (lanes 1-2) or Ad-CMV-LDLR (n = 2) (lanes 3-4) were subjected to 4% to 15% SDS-PAGE analysis under nonreducing conditions. Human LDLR expression was detected by immunoblotting, using a rabbit anti–human LDLR antibody and the ECL system. (B) LDLR–/– mice were intravenously injected with 2 × 109 plaque-forming units Ad-CMV-β-Gal (n = 5) or Ad-CMV-LDLR (n = 5). Five days after adenovirus injection, blood was drawn and plasma was analyzed for plasma cholesterol. (C) Wild-type C57BL/6J mice were intravenously injected with 2 × 109 plaque-forming units Ad-CMV-β-Gal (•) (n = 4) or Ad-CMV-LDLR (○) (n = 6). Five days after adenovirus injection, animals were intravenously injected with purified human FVIII (20 IU), and its plasma removal was monitored at indicated time points. Data represent geometric mean values and 68% confidence intervals. *P < .05, significantly different from that of control Ad-CMV-β-Gal–treated mice; Mann-Whitney U test.
