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Fig. 3. Sox8-depleted embryos fail to express Sox10 at the neural plate border. (A) Increasing amounts of Sox8mo (10 ng, 100 ng and 1000 ng) blocks translation directed by Sox8 mRNA. The same morpholino (500 ng) fails to block Sox9 and Sox10 translation. (B) Embryos injected in one blastomere at the two-cell stage with 30 ng of Sox8mo exhibit reduced Sox10 expression at stage 17, while Slug, Sox9 and Sox2 expression appears largely unaffected at this stage. (C) Quantification of the in situ hybridization results. The numbers in parenthesis indicate the number of embryos analyzed. (D) Sox8mo (500 ng) does not interfere with translation of a mutated Sox8 mRNA (mSox8). (E) Rescue experiments were performed by injection of an animal dorsal blastomere at the eight-cell stage. Bilateral Sox10 expression is rescued in Sox8mo-injected embryo by co-injection of mSox8 mRNA (Sox8mo+mSox8). Single injection of mSox8 expanded the Sox10 expression domain. Injection of a 5 bp mis-matched Sox8 morpholino (Sox8mis) had no effect on Sox10 expression. (F) Quantification of the in situ hybridization results. The numbers in parentheses indicate the number of embryos analyzed. (B,E) Dorsal view, anterior is towards the top. RNA encoding the lineage tracer β-galactosidase was co-injected to identify the injected side (red staining) (left side in B and right side in E). |
