Figure 2: Activation of the pig pluripotency network in piPSCs. PCR using human specific primers demonstrated that lentivirus delivered hPOU5F1, hSOX2, hKLF4, hC-MYC and hLIN28 human reprogramming genes were integrated into the genome of piPSCs similar to positive control hiPSCs derived using the same factors (A). Pig fibroblasts (Pig F) were negative for all 6 human genes. RT-PCR results showed that the 5 lentivirus delivered and integrated human reprogramming genes were expressed at the mRNA level in piPSCs as well as WA09 hESCs positive control cells, while pig fibroblasts were negative (B). RT negative (RT-) controls showed that there was no DNA contamination of the samples. RT-PCR with pig specific primers showed that porcine pPOU5F1, pSOX2, pNANOG and pLIN28 genes were expressed in piPSCs and therefore activation of the endogenous pig pluripotency network (C). WA09 hESC and pig fibroblasts were negative for the expression of pig specific gene expression (C).