Figure 2.

Analysis of WT male mice transduced with the AAV8 hOTC constructs

WT C57BL/6 male mice (8 weeks old) were i.v. transduced with 5.0E12 vg/kg of the indicated AAV2/8 constructs. Liver samples were collected at 14 days after viral transduction. (A) Representative WB analysis of liver extracts from control and AAV-treated animals (n = 3 per construct). Samples from two untreated mice were loaded in the first two lines of the gel. The OTC and HSP70 bands are shown. In the OTC panel, the faint band presenting a slightly faster mobility than the most prominent one corresponds to the endogenous murine OTC (see Figure S4). (B) Densitometric quantification of the Western blot (WB) of (A), relative to the OTC-WT values, normalized by the vg copy number (n = 3 per group). (C) OTC enzyme activity expressed in μmol of citrulline produced in 30 min of reaction, normalized by the vg copy number (n = 3 per group). (D) Viral genome copy number quantification by quantitative real-time PCR. The mean value of two independent determinations is indicated (n = 3 per group). Statistical significance compared to hOTC-WT is shown above the bar for each experimental group, and statistical significance between experimental groups is indicated by the horizontal line. (B) One-way ANOVA, p < 0.0001; (C) one-way ANOVA, p < 0.0001; (D) one-way ANOVA, not significant (NS); Tukey’s comp. tests, ∗p < 0.05, ∗∗p < 0.01; ∗∗∗p < 0.001.