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Fig. 7. Injection of SoxD MO into Xenopus embryos resulted in a neural defect phenotype. (A) Western blot analysis of HA-tagged proteins. SDS-PAGE was performed with protein prepared from embryos injected with 1 ng SoxD-HA(UTR+) or SoxD-HA(UTR−) mRNA either with or without 40 ng SoxD MO and cultured until stage 9. α-Actin served as an internal control. (B–D) Phenotype of embryos injected with 20 ng SoxD MO. (B) Uninjected embryo. (C) SoxD MO was injected into dorsal-animal blastomeres of eight-cell-stage embryos. (D) Linerized pCS2-SoxD-ORF (100 pg) was co-injected with SoxD MO into dorsal-animal blastomeres of eight-cell-stage embryos. (E–J) Whole-mount in situ hybridization analysis of terminal differentiation markers N-tubulin (E–H) and N-CAM (I,J). Embryos were injected with 40 ng SoxD MO into dorsal-animal blastomeres of eight-cell-stage embryos (F,H,J) and cultured until stage 15 (E,F) or 25 (G–J). (E,F) Dorsal view. |
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Fig. 2. Injection of XSIP1 MO resulted in the down-regulation of terminal neural marker genes. Whole-mount in situ hybridization analysis of N-tubulin (A,B) and N-CAM (C,D) performed on stage 28 embryos. Embryos were injected with 40 ng of either control MO or XSIP1 MO into one dorsal-animal blastomere of the eight-cell stage, and co-injected with 250 pg of nuclear-localized lacZ as a lineage tracer (red stained). (A,C) Control MO-injected embryo. Injected area (red) and marker expression (blue) overlapped. (B, D) XSIP1 MO-injected embryo. Neural markers, N-tubulin and N-CAM were not expressed in the injected area. Inset; Dorsal view of the same embryos. |
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Fig. 9. XSIP1 is required for SoxD-mediated neural induction. Embryos were injected with 500 pg SoxD mRNA alone (B,E) or co-injected with 40 ng XSIP1 MO (C,F) into the ventral marginal zone of four-cell-stage embryos and cultured until stage 32. (B,E) Ectopic neural tissue was observed on the ventral side (arrow head). (C,F) Ectopic neural tissue was not observed on the ventral side (arrow head). |