Figure 4.
Distribution of plasmid in FLR004 liver tissue. (A) Distribution maps show plasmid copy numbers per nanogram in transfected FLR004 liver lobes, which were sectioned according to an alphanumerical grid upon necropsy. The top panel shows the LLL, and the bottom panel shows the RLL. Lighter colors indicate lower copy number, and darker colors indicate higher copy number up to 445 copies per nanogram in the LLL. (B) A summary graph demonstrates comparison of average plasmid copy number per nanogram of genomic DNA among all FLR004 lobes and the untreated canine control. The caudate lobe (CL) (n = 7), RLL (n = 14), quadrate lobe (QL) (n = 4), right medial lobe (RML) (n = 10), left medial lobe (LML) (n = 8), and LLL (n = 29) are shown. Treated lobes are shown in black, and untreated lobes are shown in gray. (C) The PCR fragment amplified from genomic DNA samples isolated from a treated lobe (LLLJ1) and an untreated lobe (LMLG2) were either undigested or digested with ApaLI and analyzed by using agarose gel electrophoresis. Digestion of the expected PCR fragment (133 bp) produced 2 fragments ∼50 bp and 80 bp long. Plasmid diluted in canine genomic DNA undigested or digested with ApaLI was used as the positive control (+), and nuclease-free water digested was used as the negative control. Error bars indicate standard deviation. ****P < .0001.

Distribution of plasmid in FLR004 liver tissue. (A) Distribution maps show plasmid copy numbers per nanogram in transfected FLR004 liver lobes, which were sectioned according to an alphanumerical grid upon necropsy. The top panel shows the LLL, and the bottom panel shows the RLL. Lighter colors indicate lower copy number, and darker colors indicate higher copy number up to 445 copies per nanogram in the LLL. (B) A summary graph demonstrates comparison of average plasmid copy number per nanogram of genomic DNA among all FLR004 lobes and the untreated canine control. The caudate lobe (CL) (n = 7), RLL (n = 14), quadrate lobe (QL) (n = 4), right medial lobe (RML) (n = 10), left medial lobe (LML) (n = 8), and LLL (n = 29) are shown. Treated lobes are shown in black, and untreated lobes are shown in gray. (C) The PCR fragment amplified from genomic DNA samples isolated from a treated lobe (LLLJ1) and an untreated lobe (LMLG2) were either undigested or digested with ApaLI and analyzed by using agarose gel electrophoresis. Digestion of the expected PCR fragment (133 bp) produced 2 fragments ∼50 bp and 80 bp long. Plasmid diluted in canine genomic DNA undigested or digested with ApaLI was used as the positive control (+), and nuclease-free water digested was used as the negative control. Error bars indicate standard deviation. ****P < .0001.

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