Liver transduction efficiency by AAV6-cFVIII and AAV2-cFVIII in hemophilia A mice. (A) Southern blotting analysis. Genomic DNAs from AAV-treated mouse livers yield 2 positive bands from BamHI and BglII digestion, representing the canine FVIII light-chain (LC) and heavy-chain (HC), respectively. The standards are generated with digested plasmid DNAs spiked in liver genomic DNAs from naive hemophilia A mice. (B) RNA FISH for canine FVIII transcripts in liver sections of a mouse treated with 3 × 10¹¹  vg of AAV6-cFVIII (i-iii). (i) Section hybridized with a cocktail of canine FVIII antisense probes; (ii) same section counterstained with DAPI for nuclei; (iii) section hybridized with canine FVIII sense probes; and (iv) a naive mouse liver hybridized with canine FVIII antisense probes. Pictures were taken at 20 × magnification.