Figure 3.

Side-by-side comparison in male OTC^Spf-Ash mice transduced with AAV8-hOTC-WT and AAV8-hOTC-CO21

(A and B) 3-month-old OTC^Spf-Ash mice were i.v. injected with 2.5E11 vg/kg (n = 4), 5.0E11 vg/kg (n = 5), or 1.0E12 vg/kg (n = 5) of AAV8-hOTC-WT (A) or AAV8-hOTC-CO21 (B). Mice were sacrificed 8 weeks after viral delivery. Urine samples were collected every 2 weeks post-injection and analyzed for orotic acid. The dashed line delimits the physiological level of orotic acid in WT animals. Two-way ANOVA, 5.0E11 vg/kg, AAV8-hOTC-WT versus AAV8-hOTC-CO21, p < 0.01. (C–F) OTC protein levels were determined by WB from liver extracts from AAV8-hOTC-WT (C) and AAV8-hOTC-CO21 (D). The densitometric quantification of the OTC-specific bands, normalized by the housekeeping HSP70 protein and vg copies is shown in (E) and (F). (G) OTC enzyme activity expressed in μmol of citrulline produced in 30 min of reaction. Enzyme activity of WT and OTC^Spf-Ash liver extracts is indicated by the dashed lines. Data are shown as mean ± SEM, and statistical analyses were performed by one-way ANOVA with Turkey’s multiple comparison test (n = 3–5 per group; ∗p < 0.05). (H) The vg particles were determined by quantitative real-time PCR, and the mean values of two independent determinations are indicated (n = 4–5 per group). Data are shown as mean ± SEM, and statistical analyses were performed by two-way ANOVA with Tukey’s multiple comparison test (∗p < 0.05).