Xenbase Image ID: 78979

Fig. 2. Dishevelled regulates Notch-dependent cell fate decisions in vivo. (A-C) XDvl2 is required to regulate Notch signalling during development. Xenopus tropicalis embryos were injected at the one-cell stage with control morpholino (MOC) or one targeting Xdvl2 (MODvl2). Ciliated cell precursors were detected by α-tubulin expression (purple staining). (D) Precursors were counted within a box of standard area drawn over the centre of each embryo (see inset). Data are presented as mean number of precursors counted (±s.e.m.). Uninjected (UI) and MOC embryos were indistinguishable (A,B), whereas MODvl2 embryos exhibited a significant reduction in precursor number (C) (one-way ANOVA with Tukey's post-hoc tests). (E) qRT-PCR analysis of α-tubulin and esr1 expression in MOC and MODvl2 embryos. Expression was normalised to rpl8. Data are presented as mean fold change (±s.e.m.) in normalised expression values relative to uninjected embryos. esr1 expression was significantly increased in the MODvl2 embryos (two-tailed t-test, n=3). (F-I′) XDvl2 expression inhibits endogenous Notch signalling and rescues the NICD gain-of-function phenotype. Xenopus laevis embryos were injected in one blastomere of the two-cell embryo with mRNA encoding β-gal (F,F′), β-gal and XDvl2 (G,G′), β-gal and XNICD (H,H′), or β-gal, XNICD and XDvl2 (I,I′). (J) Ciliated cell precursors were quantified as above (see D). X-Gal staining was performed to distinguish the injected side (pale red). Images of the uninjected and injected sides of the same embryo are shown. XDvl2 promoted (G′) and XNICD inhibited (H′) ciliated cell precursor specification (two-tailed paired t-test). XDvl2 completely rescued the XNICD phenotype (I′) and there was no significant difference between the uninjected sides of any condition, and the β-gal or XNICD + XDvl2 expressing sides (two-way ANOVA and Bonferroni's post-hoc test). (K) qRT-PCR analysis of α-tubulin and esr1 expression in embryos expressing GFP or XDvl2 conducted as above (see E). esr1 expression was significantly decreased in XDvl2-expressing embryos (two-tailed t-test, n=3). Scale bars: 500 μm in A-C,F-I′. **P<0.01; ***P<0.001; ns, P>0.05. Image published in: Collu GM et al. (2012) © 2012. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license Larger Image Printer Friendly View

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