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From:
Cell Stem Cell. Author manuscript; available in PMC 2012 October 5.
Published in final edited form as:
Cell Stem Cell. 2011 February 4; 8(2): 214–227.
doi: 10.1016/j.stem.2010.12.010

Figure 6

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A β-Catenin Truncation Mutant Lacking Its TCF Transactivation Domain Sustains Pluripotency

(A) Schematic of full-length β-catenin (β-catS33A; aa 1–781) and the C-terminal deletion mutant (β-catS33AΔC; aa 1–694) used in this study.

(B) Relative protein levels of myc-tagged β-catS33A and β-catS33AΔC transgenes in mESC lines.

(C and D) TCF reporter assays (C) and qRT-PCR analysis of the β-catenin/TCF targets Axin2, Brachyury, and Cdx1 (D) demonstrate that β-catS33AΔC activates β-catenin/TCF signaling ~10-fold less than full-length β-catS33A in mESC lines.

(E) Morphology of WT-Control, β-catS33A, and β-catS33AΔC mESCs grown on gelatin-coated dishes.

(F) Expression of β-catS33AΔC in WT mESCs sustains the retention of pluripotency markers under conditions promoting differentiation (72 hr incubation).

(G) Immunofluorescent staining reveals that stable expression of β-catS33AΔC in WT mESCs inhibits the expression of the neural marker β-III-tubulin and promotes the retention of Oct-4 and Nanog in day 10 EBs.

(H) qRT-PCR analyses of several neural markers in day 10 EBs derived from the indicated control and β-catS33A overexpression mESCs demonstrates that β-catS33AΔC overexpression inhibits the expression of neural-specific genes.

(C, D, and H) Bars represent n = 3 ± SEM.

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