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. Author manuscript; available in PMC: 2016 May 18.
Published in final edited form as: Nat Cell Biol. 2015 Nov 16;17(12):1523–1535. doi: 10.1038/ncb3264

Figure 6. WNT pathway is active in naïve hESCs.

Figure 6

A: Heatmap of gene expression of WNT ligands and WNT targets in primed hESCs (H1, Elf1 AF) and naïve hESCs (Elf1). B: Wnt is activated in naïve hESCs. Endogenous Wnt signaling in naïve (Elf1) and primed (Elf1 AF) BAR-reporter cells. Scale bars represent 200µm. C: Wnt inhibitor IWP2 (2µM) and Wnt antagonist XAV939 (5µM) inhibit the reporter activity in naïve Elf1 cells after 72h. Scale bars represent 200µm. D: Wnt inhibition by IWP2 (2µM, 48h) decreases OCR changes after FCCP in naïve hESCs (Elf1, WIN1) and in naïve hESCs transitioning to primed (WIN1 AF). A trace of OCR changes is presented in Elf1 (n=8 for Elf1, n=6 for Elf1+IWP2; s.e.m.). OCR changes after FCCP were quantified (n=8 for Elf1, n=6 for Elf1+IWP2, WIN1, WIN1+IWP2, WIN1AF, n=7 for WIN1AF+IWP2; s.e.m.; p=0.0009 for Elf1+IWP2 vs. Elf1, p=0.0084 for WIN1+IWP2 vs. WIN1, p=0.0006 for WIN1AF+IWP2 vs. WIN1AF; 2-tailed t-test). E: Wnt inhibition by IWP2 (2µM, 72h) downregulates NNMT and miR-372 expression in naïve hESCs (Elf1) as shown by qPCR analysis. (n=3; s.e.m.; p=0.04 for miR-372, p=6.44E-06 for NNMT; 1-tailed t-test). F: Model of self-reinforcing loop between WNT and NNMT in primed hESCs. For raw data, see Supplementary Table 4. n= number of biological replicates.