A, Tissue sampling scheme of pig liver across different lobes: right lateral lobe (RLL), right medial lobe (RML), left medial lobe (LML), left lateral lobe (LLL), and quadrate (Q) lobe. Proximal and distal samples to the site of catheter injection in the CHD were taken among all lobes. Pig no. 3 liver is depicted in A, harvested 1 week after injection for subsequent tissue analysis. B, PCR on genomic DNA revealed the expected band (550 bp) for hFIX DNA in all liver lobes tested. C, RT-PCR was performed on RNA extracted at 1 week after injection demonstrated the expected band size. RNA extracted from HepG2 cells transfected with pT-LP1-hFIX plasmid serves as one positive control. For both PCRs, positive control (CTL) is hFIX plasmid, and negative control is non-injected pig liver tissue. D, Western blot on liver tissue demonstrated the correct size of hFIX (70kDa) with a low-level cross reactivity for porcine FIX at the same molecular weight. Quantification of hFIX expression by western blot band intensity and normalized with beta-actin is provided. (CTL = control, non-injected pig liver).