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. Author manuscript; available in PMC: 2016 Mar 25.
Published in final edited form as: Cell Rep. 2016 Mar 3;14(10):2301–2312. doi: 10.1016/j.celrep.2016.02.036

Figure 1. YAP Overexpression Promotes the Generation of Human Naive ESCs.

Figure 1

(A) YAP protein is localized to the nucleus in both the trophectoderm and the ICM of human blastocysts, as shown by immunofluorescence. White, DAPI; green, OCT4 (ICM); yellow, CDX2 (trophectoderm); red, YAP. Representative image of n = 16 human blastocysts. Scale bar, 20 μm.

(B) In naive medium N2B27+2iFL, human H9 ESCs with YAP overexpression (H9-YAP) maintain expression of the pluripotency marker AP, while control H9 cells differentiate and do not express AP. Scale bar, 500 μm.

(C) H9-YAP cells in N2B27+2iFL have a naive-specific dome-like colony morphology and show strong positive immunostaining for pluripotency markers SSEA3, SSEA4, TRA-1-60, TRA-1-81, OCT4, and NANOG. Black scale bar, 500 mm; white scale bar, 150 μm.

(D) Growth curves of H9 in DF12+bFGF primed medium, H9-YAP in DF12+bFGF primed medium, and H9-YAP in N2B27+2iFL naive medium after trypsinization to single cells. Only H9-YAP cells in naive medium have a high proliferate rate. Error bars represent SD.

(E) H9-YAP cells in N2B27+2iFL have a normal female karyotype (46, XX), as evaluated after 20 passages in naive culture conditions.

(F) H9-YAP cells in N2B27+2iFL are able to form teratomas comprising tissues derived from all three germ layers. (a) Adipocytes (mesoderm). (b) Neural tissue (ectoderm). (c) Epithelium (endoderm). White scale bar, 200 μm; black scale bar, 50 μm.