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Experiment details for ebf2

Logan MA et al. (2005) Assay

Identification of shared transcriptional targets for the proneural bHLH factors Xath5 and XNeuroD.

Gene Clone Species Stages Anatomy
ebf2.S laevis NF stage 18 neuroectoderm , nervous system , neural tube

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  Fig. 2. Xath5 or XNeuroD overexpression induces ectopic expression of candidate target genes identified by FDD. Xath5 or XNeuroD mRNA was injected into 1 cell of 2-cell stage embryos with GFP mRNA as an injection marker. Whole mount in situ hybridization was performed to visualize expression of candidate target genes. Xath5 overexpression induces significant ectopic broad expression of elrC (A) and XETOR (B) on the injected side of the embryo. The novel gene SBT1 is expressed in the developing nervous system (arrows in panels C and J) and displays a scattered pattern of ectopic expression on the Xath5-injected side. Xath5 overexpression results in upregulation of Xgadd45γ (D), Xebf2 (E) and Xebf3 (F) within the developing nervous system, with weaker ectopic expression also induced in lateral ectodermal regions. At early tailbud stages, Xath5 induces ectopic expression of NKL (G) and SVOP (H) on the injected side of the embryo. NeuroD overexpression also induces broad robust ectopic elrC expression (I) and scattered ectopic expression of SBT1 (J). At early tailbud stages, NeuroD injection promotes scattered ectopic expression of NKL (K) and SVOP (L) on the injected side. Ectopic gene expression is indicated with brackets. Embryos in panels (A) and (I) are presented as dorsal views with anterior to the top and injected side on the left. Tailbud stage embryos in panels (H) and (L) are shown as lateral views with anterior to the right.

Gene Clone Species Stages Anatomy
ebf2.S laevis NF stage 18 neuroectoderm , neuron , neural tube

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  Fig. 4. Dominant-negative Xath5 inhibits expression of Xath5/XNeuroD shared target genes. Injection of Xath5EnR inhibits endogenous expression of elrC (A), SBT1 (B), Xebf2 (C), XETOR (D), Xebf3 (E), SVOP (F) and Xgadd45γ (G) in mid-neurula and tailbud stage embryos. Xath5EnR injection also blocks endogenous expression of NKL (H) at tailbud stages. Endogenous expression of target genes is indicated by arrowheads, and loss of or reduced expression of genes on Xath5EnR injected side is indicated by brackets. All embryos are shown as dorsal views with anterior to the top and Xath5EnR-injected side on the left, except for the NKL probed embryo (H), which is shown as a lateral view with anterior to the right. Pink staining (A) and light blue staining (B) correspond to β-galactosidase injection tracer. Embryos in remaining panels (C) were presorted for GFP to identify injected side. Abbreviations: sc, spinal cord; e, eye.