Figure 1: EpEX and/or EpCAM promote the reprogramming of Oct4-GFP mouse embryonic fibroblasts (MEFs) into iPSCs. | Scientific Reports

Figure 1: EpEX and/or EpCAM promote the reprogramming of Oct4-GFP mouse embryonic fibroblasts (MEFs) into iPSCs.

From: EpEX/EpCAM and Oct4 or Klf4 alone are sufficient to generate induced pluripotent stem cells through STAT3 and HIF2α

Figure 1

(A) The time line of iPSC induction. The expression of exogenous reprogramming genes was induced by doxycycline from day 2–21. After induction, GFP-positive colonies were formed with either OSKM (O: Oct4; S: Sox2; K: Klf4; M: c-Myc) or OSKME (OSKM + EpCAM; E: EpCAM), with or without EpEX (soluble extracellular domain of EpCAM) treatment. (B) Fluorescence microscopy was used to reveal Oct4 expression in Oct-GFP iPSC colonies on day 25. Scale bar: 50 μm. (C) The formations of Oct4-GFP iPSCs was evaluated by examining colony numbers on day 25 under fluorescence microscopy. (D) Alkaline phosphatase staining was performed to examine undifferentiated iPSCs. Scale bar: 50 μm. (E) Expression of pluripotency genes in iPSCs. Q-PCR analysis was used to show that iPSCs express the endogenous pluripotency genes Oct4, Sox2, and Nanog. Parental MEFs were used as a control (n = 3). (F) Western blot analysis was performed to reveal that the iPSCs express Oct4, Sox2, and Nanog protein (n = 3). (G) The pluripotency of iPSCs was examined by teratoma formation. Mice were injected with OSKM and OSKME + EpEX iPSCs (n = 3). Scale bar: 50 μm.

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