Expression of Stabilized β-Catenin in WT mESCs Inhibits Neural Differentiation and Prolongs the Retention of Pluripotency
(A) Morphology of WT mESCs expressing stabilized β-catenin (WT-β-catS33A, independent clones A3 and B2), relative to control mESCs stably transfected with empty vector (WT-Control) and DKO mESCs. Scale bar represents 100 μm.
(B) Immunofluorescent staining of day 14 EBs reveals that β-catenin-overexpressing EBs fail to generate neural tissue, as determined by staining for the neural marker β-III-tubulin. Scale bar represents 20 μm.
(C) qRT-PCR analyses of several neural markers in day 14 EBs derived from control and β-catenin-overexpression mESCs demonstrates that β-catenin overexpression inhibits the expression of neural-specific genes. Bars represent n = 3 ± SEM.
(D) Teratomas derived from β-catenin-overexpressing mESCs failed to generate neural tissue in vivo, as determined by immunohistochemical staining for β-III-tubulin. Scale bar represents 500 μm.
(E) β-catenin overexpression prolongs the retention of the pluripotency markers (Oct-4, Nanog, and Sox2) under conditions promoting differentiation (72 hr incubation).