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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. |