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Fig. 4. Apoc1 is required for neural border formation and neural crest emergence. (A) Whole mount in situ hybridization for the neural crest (NC) markers c-myc, sox9, snai2, twist1, id3. Control MO injected embryos are shown in upper panel and Apoc1 tbMO injected ones are shown in lower panel. (B) The reduction of sox9 and snai2 expression by the Apoc1 tbMO could be rescued by coinjection of 400 pg of apoc1 mRNA (upper panel) but not by coinjection of 400 pg of sp-apoc1 mRNA (lower panel). (C) Whole mount in situ hybridization for the neural plate border specifiers msx1, pax3, zic1, the neural marker sox2, and the non-neural ectoderm marker xk81a1. Control MO injected embryos are shown in upper panel and Apoc1 tbMO injected ones are shown in lower panel. One of the dorsal blastomeres was injected with 30 ng of either control MO (upper panels in A, C) or Apoc1 tbMO (lower panels in A, C) together with 100 pg lacZ mRNA. Embryos were fixed at stage 15. The injected side was traced by X-gal staining (blue). |
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Fig. 6. Apoc1 and gbx2.2 are required for emergence of neural crest cells, but are independently regulated by Wnt signaling. (A-H) The expression of apoc1 and gbx2.2 is independently regulated, but both genes are required for neural crest induction. Apoc1 expression in Gbx2.2 (A) or control MO (B) injected embryos and gbx2.2 expression in Apoc1 (C) or control MO (D) injected embryos. The reduction of sox9 and snai2 expression was not rescued by coinjection of 100 pg gbx2.2 mRNA in Apoc1 (E,G) or 400 pg apoc1 mRNA in Gbx2.2 (F, H) depleted embryos. 30 ng of MO and 100 pg lacZ mRNA was injected into one side of a single dorsoanimal blastomere of 8-cell stage embryos. The embryos were fixed at stage 15. (I-R) Apoc1 and gbx2.2 are both required for Wnt-mediated neural crest induction in vivo. 200 pg of GR-ctnnb1 and 100 pg lacZ mRNA were injected into one side of a dorsoanimal blastomere of 8-cell stage embryos and the embryos were treated with DEX at stage 10.5. The embryos were fixed at stage 15 for whole mount in situ hybridization with snail2 (I), sox9 (J), apoc1 (K), gbx2.2 (L), msx1 (M), and pax3 (N). Ectopic expression of gbx2.2 in GR-ctnnb1 injected embryos was not reduced by coinjecting 30 ng Apoc1 tbMO (P), but the anterior expansion of snai2 was rescued (O). Ectopic expression of apoc1 in GR-ctnnb1 injected embryos was not reduced by coinjecting 30 ng Gbx2.2 MO (R), but the anterior expansion of snai2 was rescued (Q). Blue arrowhead indicates ectopic expression. |