Figure 4.

The dual vector system leads to specific expression of the recombined transgene. (A) Schematic representation of the experimental animal setting to assess the specific expression of the reconstituted transgene and absence of truncated aberrant dystrophin isoforms coming from the single AAV vectors. (B) Scheme of the epitopes recognized by the two dystrophin antibodies used. Dys B antibody recognizes the N terminal part of dystrophin, while Dys 2 recognizes the C terminal part. (C) Dystrophin Western Blot results on TA and DIA of mice injected with 5′ and 3′ vectors singly or together and with PBS as a control. Dystrophin protein levels are normalized for GAPDH expression. The hypothetical truncated protein product originating from the 5′ vector would have a molecular weight around 151 kDa (red square), while the hypothetical protein originating from the 3′ vector would have a molecular weight of 140 kDa (blue square). (D) RTqPCR analysis performed on RNA extracted from TA and DIA muscles using primers to specifically only the 5′ or 3′ transgene and the recombined transgene (different colors are associated with different groups).