Induced pluripotent stem cells (iPSCs) generated from mouse embryonic fibroblast (MEF) cells of wild-type mice and Pax6 mutants exhibit mouse embryonic stem (mES) cell characteristics. (A) Morphological features of iPS (upper line) colonies and their alkaline phosphatase staining (lower line), showing mES-cell-like morphological features distinct from those of MEFs at 5 days after Dox-inducible lentiviral transduction. Scale bar=100 μm. (B) Immunofluorescence staining demonstrates that iPS clones are positive for Nanog and SSEA-1. Nanog scale bar=50 μm, SSEA-1 scale bar=100 μm. (C) Hierarchical cluster analysis of global gene expression microarray data from iPSCs of Wt, Hetz, Homo, mES, and MEF cells. (D) Scatter plot comparing global gene expression profiles among Wt, Hetz, Homo, mES, and MEF cells. (E) Bisulfite genomic sequencing of the promoter regions of Oct4 and Nanog.
Open and closed circles indicate non-methylated and methylated CpGs, respectively. (F) Histological sections show differentiation into all three germ layers: neural rosettes (ectoderm), endothelium (endoderm), and muscle (mesoderm). Black arrows heading to typical tissues. Scale bar=100 μm.