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Figure 4. MarvelD3 is required for neural crest formation. Neural crest induction was analysed at stage 15 by WISH using probes against the neural crest markers twist, snai1, foxd3 and snai2/slug, respectively, in control morpholino (A–D), MD3AB morpholinos (E–H) and embryos co-injected with MD3AB morpholino + 7mut-marveld3 RNA (I–L). The number of embryos analysed is indicated at the bottom right of each panel; red asterisk, injected side of the embryo with the light blue β-galactosidase staining; purple corresponds to the WISH staining; scale bar, 500 µm. (M) Quantification of neural crest induction was performed for twist, snai1, foxd3 and snai2/slug. ANOVA values are provided in parenthesis in the title of each graph. Student t-test p values and the number of embryos analysed are indicated under the graph. See Supplemental Fig. S1 for images of embryos injected with individual MD3A or MD3B morpholinos and corresponding rescue experiments. |
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Supplemental Figure S1. Depletion of MarvelD3 and neural crest development. (A) gfp-fl marveld3 was inserted in pCS2+ vector (BamH1/EcoR1). 7mut-marveld3 was subcloned in pCS2+ vector (BamH1/ EcoR1). Restriction sites: underlined; mutated bases of marveld3: underlined lower cases. (B) neural crest induction (stage 15) was determined by WISH for twist, snai1, foxd3 and snai2/slug in control morpholino- (i), MD3A morpholino- (ii), MD3B morpholino-injected embryos (iv) and embryos co-injected with FL marveld3 (iii) or 7mut-marveld3 mRNA (v), respectively; the number of embryos analysed is indicated at the bottom right of each panel; red asterisk, injected side of the embryo with light blue β- galactosidase staining; purple corresponds to the WISH staining; scale bar, 500 μm. |
