Additional supporting information available online.

STEM_281_sm_suppinfo.doc91KSupplemental Info
STEM_281_sm_suppinfoFigureS1.tif2003KSupplemental online Figure 1. Generation of iPS cells from DP cells with three reprogramming factors. FACS sorted DP cells were 2x infected with retroviruses expressing Oct4, c-Myc, and Klf4 (3TF) and no Sox2. A typical Oct4-GFP positive iPS colony reporting activation of endogenous Oct4 expression is shown (right). The Phase image shows the typical ES/iPS morphology (left).
STEM_281_sm_suppinfoFigureS2.tif2663KSupplemental online Figure S2. Immunofluorescence staining of pluripotency markers in DP-derived 4TF iPS cells. 4TF iPS cells were stained with antibodies against Nanog, Oct4, Sox2, and SSEA-1 (red). Nuclei were visualized by DAPI (blue).
STEM_281_sm_suppinfoFigureS3.tif5985KSupplemental online Figure S3. Integration analysis of the four-transgene factors by Southern blotting. Genomic DNA isolated from MEFs, DP cells, 2TF-1, 2TF-10 and 4TF-1 iPS cells were digested with Bgl II, and hybridized with Oct4, Klf4, Sox2 and c-Myc cDNA probes, respectively. Red stars indicate the integration of 1-4 four copies of the exogenous reprogramming genes. The blue triangle denotes the endogenous gene.

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