Additional Supporting Information may be found in the online version of this article.

sc-12-0316_sm_SupplFigure1.pdf333KSupplementary Figure S1: Oct4-MerCreMer transgene accurately reflects endogenous Oct4 expression (A, B): Southern blot indicating correct targeting at the modified Oct4 locus. (C-F): Correlation of EGFP with endogenous Oct4 expression. Oct4-CreMerCre mT/mG fibroblasts were reprogrammed in the absence of Tamoxifen for 20 days before addition of Tamoxifen for 16 hours. Colonies were dispersed into single cells (C): before sorting (D, E): after sorting by FACS (F). (G): RT-PCR analysis shows that endogenous Oct4 gene expression is restricted to cells expressing EGFP. (H): Genomic PCR detects all 4 reprogramming viruses are integrated in both tdTomato and EGFP cell populations.
sc-12-0316_sm_SupplFigure2.pdf456KSupplementary Figure S2: Derivation and characterization of iPS cell lines from Oct4- MerCreMer; mT/mG embryonic fibroblasts reprogrammed in the absence of tamoxifen. (A): 3F-10, an iPS cell line derived after 3F OSK reprogramming, phase image. (B): 3F-10, SSEA1 staining. (C): 3F-10, tdTomato expression. (D): 3F-10, Nanog staining. (E): 7-8-8, an iPS cell line derived after 4F OSKM reprogramming, phase image. (F): 7-8-8, Oct4 staining. (G): 7-8-8, tdTomato expression. (H): 7-8-8, Nanog staining. These lines exhibit a very low background of tamoxifen independent EGFP activation common to post-translational inducible Cre systems (I): RT-PCR analysis of iPS cell lines 3F-10, 7-8-8 and 7-8-10. (J) In vitro differentiation potential during embryoid body formation. (K): Demethylation of Oct4 and Nanog promoters confirmed by bisulfite treatment and sequencing. (L-N): In vivo pluripotency determined in teratoma derived from 3F10 iPS cells, from left to right, neural tissue, muscle, and intestine-like epithelial tissue with goblet cells.
sc-12-0316_sm_SupplMethods.pdf115KSupplementary Data
sc-12-0316_sm_SupplTable1.pdf12KTable S1: Primers used in this study

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