Southern and RT-QPCR analysis of transduced livers. Liver DNA from Hem-A/Rag1 mice was extracted 18 weeks after portal vein injection with 3 × 10¹¹ vg AAV-hFVIII-HC and AAV-hFVIII-LC (samples 1.1, 1.2, 2.0, 2.1, 2.2, 3.0), AAV-hFVIII-LC (sample 3.2), or AAV-hFVIII-HC (sample 4.2). For the hFVIII-HC copy number determination, the DNA (10 μg) was digested with HindIII (A), while for the light chain copy number the DNA was digested with PstI (B). ³²P-random primer labeled heavy- and light-chain probes were used for the respective blots. Heavy- and light-chain RNA levels were determined by RT-QPCR using heavy- and light-chain–specific primers and probes. The DNA copy numbers are expressed as copies per diploid genome. The RNA levels are expressed as copies/10 ng RNA and have been normalized to 1 DNA copy/cell by dividing by the DNA copy number that was determined by Southern blot analysis.