FIG. 5.

rAAV-mediated gene expression and quantification of rAAV genomes in partially hepatectomized mice. (A) Plasma hF.IX levels of 20 mice injected with AAV-EF1α-F.IX at a dose of 2.4 × 10¹¹ vg per mouse. Half of the mice underwent partial hepatectomy (PHx) 12 weeks postinjection. Vertical bars indicate standard deviations. (B) Southern blot analysis of liver DNAs from the mice in panel A. All animals were sacrificed 18 weeks postinjection (6 weeks after partial hepatectomy for the partial hepatectomy group), and 20 μg of total genomic liver DNA was subjected to Southern blot analysis with BglII digestion and hybridized to a BglII F.IX probe (28). Copy number standards are indicated as 0.0 to 6.0 copies/cell. The numbers above each lane indicate individual mouse numbers. Ethidium bromide staining of the gel showed that all the lanes had the same amount of digested DNA (data not shown). Both blots were from the same membrane. (C) Comparison of rAAV vector genome copy numbers per cell before and after partial hepatectomy. The intensity of each band in panel B was determined by densitometry. A Student t test revealed a statistical difference between “Pre PHx” and “Post PHx” values (P < 0.0002). Vertical bars indicate standard deviations.