Summary of porcine tumor growth in vivo.

<p>^a Offspring sired by clone 63–3</p><p>^bIM, intramuscular; SQ, subcutaneous; IT, intra-testicular</p><p>^cConcentration of 2x10⁹ pfu AdCre/ml</p><p>^dSize 10 days post-injection (cm)</p><p>^eSize 20 days post-injection (cm)</p><p>^fRatio of normalized RNAseq levels between AdCre induced tumor tissue RNA/untreated transgenic muscle tissue RNA</p><p>Summary of porcine tumor growth in vivo.</p

Inducible expression of <i>KRAS</i>^<i>G12D</i> and <i>TP53</i>^<i>R167H</i> is transforming and tumorigenic.

<p>(<i>a</i>) RT-PCR analysis of <i>KRAS</i>^<i>G12D</i> and <i>TP53</i>^<i>R167H</i> mRNA expression in the fibroblast cell lines from each of the 4 transgenic clones treated with AdCre or AdGFP. (<i>b</i>) Comparison of cell morphology between AdCre and untreated control cells in culture, stained with H&E. (<i>c</i>) Normalized MFU measured by FACS at time points following Carboxyfluorescein succinimidyl ester (CFSE) dye loading of cells. (<i>d</i>) Graphical analysis of the mean number of migrating cells from triplicate plating of each of the 4 cell lines. (<i>e</i>) Graphical analysis of the mean number of colonies growing in soft agar for each cell line from triplicate plating. (<i>c-e</i>: all data points are the mean of the 4 cell lines derived from pigs 63–1, 63–2, 63–3, 63–4; error bars = SD; *p-value ≤ 0.05; **p-value ≤ 0.01).</p

Injection of AdGFP into transgenic pigs did not induce tumors.

<p>The lack of any detectable changes to the injection site on d20 post- injection is shown at the surface (a–c); in underlying tissue (d–f); or upon histological examination (g–i).</p

Tumors arose from each AdCre treated cell line injected into immune compromised mice.

<p>Similar tumors (a) developed from all AdCre cell lines and no tumors developed from AdGFP cell injections; Histological analysis (b) revealed the tumors to be densely cellular non encapsulated and infiltrative neoplasm. 10x and insert 40X H&E Stain.</p

Inducible expression of <i>KRAS</i>^<i>G12D</i> and <i>TP53</i>^<i>R167H</i> is tumorigenic in transgenic pigs.

<p>(<i>a–c</i>) Ultrasound images of tumors developing 10 days after, and (<i>d–f</i>) images of these tumors at necropsy 20 days after intramuscular (IM, Pig 1), subcutaneous (SQ, Pig 3), or intra-testicular (IT, Pig 1) injection of AdCre in transgenic oncopigs containing a Cre-inducible vector encoding <i>KRAS</i>^<i>G12D</i> and <i>TP53</i>^<i>R167H</i>; (g–i) H&E stained sections show the tumor and adjacent normal tissue (2x) with an inserted high magnification photo (20x).</p

The Cre-inducible vector encoding <i>KRAS</i>^<i>G12D</i> and <i>TP53</i>^<i>R167H</i>.

<p>(<i>a</i>) Schematic diagram of the vector encoding Cre-inducible <i>KRAS</i>^<i>G12D</i> and <i>TP53</i>^<i>R167H</i>. (<i>b</i>) PCR analysis for the presence of <i>KRAS</i>^<i>G12D</i> and <i>TP53</i>^<i>R167H</i> in genomic DNA isolated from the indicated cloned offspring.</p