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Supplementary material Fig. S5. Flop1/2 also regulate head development. (A) Flops-overexpressing embryos with mild gastrulation defects showed micro- or anencephalic phenotypes. Flop1 or Flop2 mRNAs were injected into the dorsal side of four-cell-stage embryos. (B–D) Spatially confined expression of Flops resulted in the development of microcephaly. (B) Flops mRNAs were injected into two dorso–animal blastomeres at the 16-cell-stage to avoid generating spina bifida. (C) Injected embryos showed normal gastrulation but exhibited microcephaly and the reduction of body pigmentation (arrows). (D) Quantified data. n: Number of examined embryos. Chi-square test, *P<0.05 and ***P<0.001. ns: No significance. (E, F) Formation of the anterior axial mesoderm and neural crest were defective in Flops-injected embryos. (E) Embryos were prepared as in B. Expression of dorsal midline marker shh was attenuated in Flops-injected region (arrow). (F) mRNAs were injected into one dorsal blastomere at the four-cell-stage. Expression of the neural crest marker slug was diminished in Flops-injected region (arrow). The injected region was visualized by immunostaining of the co-injected tracer GFP (150 pg) using an anti-GFP antibody after WISH. |
