Supplementary Figure 3: Schematic overview for the blastocyst complementation studies using the porcine model.

The figure demonstrates the flow of experiments that were performed, which began with the generation of ETV2 mutated porcine fibroblasts using CRISPR/Cas9 gene-editing technology. Somatic cell nuclear transfer (SCNT) technology was used to clone the ETV2 mutated nucleus from fibroblasts into the enucleated pig primary oocytes. The developing morulae from the cloned oocytes were injected with either porcine stem cells (blastomeres) for pig-pig or human hiPSCs for human-pig complementation studies. The chimeric complemented embryos were either cultured in vitro or transferred into synchronized gilts for in vivo studies. We performed proliferation and survival assays, immunohistochemistry, in situ hybridization, human-pig cell integration and communication studies and TUNEL assays in vitro. For in vivo studies, embryos were harvested at defined embryonic stages [E18, E24 or FT (full term)] and analyzed using FACS, gDNA and RNA qPCR, immunohistochemistry, in situ hybridization or methylcellulose assays.