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Fig. 2. Dysregulation of Custos inhibits the canonical Wnt pathway. (A and B) Overexpression or depletion of Custos reduced organizer gene expression (A) and anterior markers (B). Custos RNA (1.5 ng) or Custos-MO (50 ng) was injected with LacZ (250 pg) RNA into one dorsal cell of four-cell embryos. At stage 10.5, the expression level of organizer genes Xnr3 and Goosecoid (Gsc), or at stage 26, the expression of anterior markers Rx1, N-CAM and Wnt1, was examined by in situ hybridization. Red staining indicates the injected side. The number of embryos scored is shown on each panel. (C) Custos regulates nuclear accumulation of β-catenin at stage 10.5. Custos-MO (50 ng) or myc-Custos RNA (1 ng) was injected into one dorsal blastomere of four-cell embryos. mCherry RNA (250pg) was coinjected and red fluorescence was used as tracer for the injected side. Embryos were fixed at stage 10.5, and immunohistochemistry was performed with anti–β-catenin and anti-myc antibodies. Arrowheads indicate cells with nuclear accumulation of β-catenin. |
