Figure 2.

U1+5a-mediated rescue of hFVII expression by hydrodynamic injection. (A) hFVII levels in plasma (mean and 95% confidence intervals) at 48 h upon hydrodynamic injection in mice (four mice per group) of two doses of pFVII plasmids (1 or 2 μg g⁻¹ mouse body weight; 1× or 2×, respectively) without or with a corresponding 1.5× molar excess of pU1+5a. The negative control (no plasmid injection) is represented by mice injected with PBS only. Inset: hFVII levels in plasma (mean and 95% confidence intervals) at 48 h upon hydrodynamic injection in mice (five mice per group) of 2 μg g⁻¹ mouse body weight of pFVII plasmids with a corresponding 1.5× molar excess of pU1wt. (B) Analysis of hFVII mRNA forms in mouse livers. The aberrant (a) and normal (n) transcripts are depicted on the right. Arrows indicate primers used for RT-PCR. Amplicons were resolved on a denaturing capillary electrophoresis system. Histograms report the quantification of transcripts, expressed as percentage (mean and 95% confidence intervals) of total hFVII forms, in liver mRNA from mice injected with pFVIIwt or pFVII+5A, without or with a 1.5× molar excess of pU1+5a. M, size marker; bp, base pairs. (C) Representative examples of liver sections stained with a species-specific anti-hFVII antibody (green). Mice were injected with 2 μg g⁻¹ mouse body weight of plasmids pFVIIwt or pFVII+5A, without or with a 1.5× molar excess of pU1+5a. Images are taken at 100× magnification. Scale bar, 50 μm.