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. 2015 Apr 23;33(5):1390–1404. doi: 10.1002/stem.1926

Figure 2.

Figure 2

Efficient reprogramming of Rex1‐GFP MEFs to integration‐free iPSCs in N2B27/leukaemia inhibitory factor by 6F. (A): AP+ colonies of expressing 4F or 6F from the episomal vectors. Left panel: AP+ colonies. Right panel: colony numbers. **, p < .01. (B): A 6F GFP+ colony from the episomal vectors. Scale bar = 200 µm. (C): Characterization of iPSCs for loss of the episomal vectors. Whole cell lysis was used to detect residual episome or genome‐integrated episomal vectors (top four panels). Actb was used as a genomic DNA polymerase chain reaction (PCR) control. The bottom four panels show RT‐PCR‐amplified transcripts of exogenous factors. Actb expression was used as the control. E‐iPSCs were produced using episomal vectors. PB‐iPSCs were reprogrammed using piggyBac vectors. (D): Quantitative real‐time PCR analysis of pluripotent gene expression in iPSCs from 6F‐episomal vectors. The expression is shown relative to Gapdh and normalized to ESCs. (E): Teratomas derived from 6F E‐iPSCs. (F): Chimeric mice from iPSC injection into host blastocysts. Experiments were repeated at least three times, and the error bars show standard deviations from the mean of triplicate determinations of one representative experiment. Abbreviations: AP, alkaline phosphatase; ESC, embryonic stem cell; GFP, green fluorescent protein; iPSC, induced pluripotent stem cell; MEF, mouse embryonic fibroblast cell.