FIG. 3.

Quantitative examination of the flag-tagged HH tri-AAV vectors in mdx4cv mouse muscle. (A) Schematic outline of the flag-tagged HH tri-AAV vectors. The structure is identical to the original HH tri-AAV vectors except the addition of a flag tag at the C-terminal end of the human dystrophin coding sequence. Locations of the primers used for quantitative RT-PCR are marked (see Table 3 for details). (B) Representative Pan-Dys and flag antibody immunofluorescence staining photomicrographs. Top two panels show a revertant myofiber (positive for Pan-Dys staining but negative for flag staining). *The same myofiber in serial sections. (C) Quantification of human dystrophin reconstitution in mdx4cv mice infected by one, two, or all three vectors of the flag-tagged HH tri-AAV vectors. N=14 for WL30/WL33/WL38 coinfected mice; N=6 for WL30 infected mice; N=8 for WL30/WL38 coinfected mice; N=8 for WL38 infected mice. *Significantly higher than other groups by flag antibody staining. ^#Significantly higher than other groups by Pan-Dys antibody staining. Cross, significantly higher than that of flag antibody staining in the same group. (D) Quantitative RT-PCR results. The primer set used in each qRT-PCR is indicated in the x-axis. N=14 for tri-AAV infected mice; N=6 for noninfected control mice. *Significantly higher than that of uninfected. (E) Representative flag, Pan-Dys, and Hum-Dys antibody immunofluorescence staining photomicrographs. *The same myofiber in serial sections. (F) Quantification of human dystrophin reconstitution in mdx4cv mice infected with high-dose flag-tagged HH tri-AAV vectors. N=2 mice for each group. The error bar stands for the difference between the mean value and each individual value. RT-PCR, reverse transcription polymerase chain reaction.