Postneonatal Mortality and Liver Changes in Cloned Pigs Associated with Human Tumor Necrosis Factor Receptor I-Fc and Human Heme Oxygenase-1 Overexpression

Figure 2

Copy number, hHO-1 and shTNFRI-Fc expression level of transgenic pigs. (a) While a C4-shTNFRI-Fc/hHO-1 piglet have at least 4 copies in genome, all C15-shTNFRI-Fc/hHO-1 piglets harbor 15 copies of the hHO1 and shTNFRI-Fc transgene in their genome. Genomic DNA from transgenic pig was extracted and analyzed using gradient PCR. (b) Relative expression of HO-1 mRNA in liver was detected by real-time PCR. The experiment is performed in 9 replications and data are presented as mean values ± standard deviation. Different superscript means significance (). (c) Quantitative analysis of shTNFRI-Fc analysis in liver was performed using ELISA. The experiment is performed in eight replications and data are presented as mean values ± standard deviation. (d) HO-1 in the liver protein lysate of transgenic pigs. Relative expression of HO-1 protein in liver of transgenic pigs was detected by Western blot. Anti-β-actin staining was used as a loading control. A clear overexpression of shTNFRI-Fc/hHAHO1/B KO compared to shTNFRI-Fc/hHAHO1/A KO is evident in all transgenic pigs.

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