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Fig. 5
Foxo3b gain-of-function inhibits Wnt/β-catenin signaling in embryos.
(A) Foxo3b inhibited β-catenin/T cell factor activity in embryos. A1, 1-cell stage embryos were injected with a mixture of plasmids as indicated, together with pFR-luc as a reporter gene and pTK-renilla as an internal control; luciferase activity was measured after 11h. Date presented are the average (±SEM) of four independent experiments. A2, 1-cell stage embryos were injected with 3xTOPFlash, and the plasmids as indicated, together with pTK-renilla as an internal control; luciferase activity was measured after 11 h, performed in triplicate. "**" indicates p<0.01; "***" indicates p<0.001. (B) Gain-of-function of foxo3b resulted in suppression of Wnt/β-catenin signaling in embryos. B1-B2, foxo3b over-expressed embryos showed reduced vox expression (arrowheads in B1 and B2) compared to wild-type. B3-B6, the expression of ventral marker vent and ved (domain width indicated by arrowheads) decreased in 70% (n = 20) and 75% (n = 16) of foxo3b-ATG-MO-injected embryos respectively. B7-B10, the expression domain of dorsal marker gsc expanded in most foxo3b over-expressed embryos. Embryos were injected with 2 ng GFP mRNA (control) or 2 ng foxo3b mRNA. B1-B6, animal pole views with dorsal to the right; B7-B8, dorsal views with anterior on top; B1-B8, shield stage.