Figure 5.

Transgene mRNA oxidation after rAAV injection in dystrophic mice and dogs. (a) RNA Immunoprecipitation analysis of GFP mRNA oxidation at 60 days postinjection in the TA muscles of mdx4Cv mice and C57BL/6J (B6) controls, using a specific anti-8-HOG antibody (15A3). No background signal was obtained with the isotype control (IgG CTL). The GFP mRNA copy number was measured by RT-qPCR after Immunoprecipitation and expressed as a percentage of INPUT. The data represent the median value (central line), the first and third quartiles (box) and first and ninth deciles (whiskers) of four (B6) or seven (mdx4Cv) mice per group. The shaded area delineates qPCR limit of detection. Statistics: Mann–Whitney’s U-test; **P < 0.01. (b) Microdystrophin (µDys) mRNA oxidation after rAAV8 locoregional injection in Golden Retriever muscular dystrophy (GRMD) dogs. % INPUT were obtained with the same protocol as in a and plotted as a function of µDys RQ, measured by RT-qPCR and normalized with the endogenous target Rpl32. The 19 samples represent various forelimb muscles collected from four GRMD dogs injected with a similar vector dose (1 × 10¹³ vg/kg) and euthanized between 13 and 16 weeks postinjection. Statistics: Mann–Whitney’s U-test between two distinct categories of samples (RQ < 1 versus RQ > 2); **P < 0.01.