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Fig. 4. Analysis with antisense morpholino oligonucleotides against Wise. (A) Antisense morpholino oligonucleotides against Xenopus Wise specifically blocks translation of Xenopus Wise RNA. Embryos were injected as indicated, animal caps were cut at stage 8, and the caps were collected at stage 9. Cell extracts were analysed by western blotting using anti-Flag and anti-HSP70 antibodies. (B-E) Phenotypes arising from whole-embryo injection of control (B,C) or Wise antisense (D,E) morpholino oligonucleotides at stage 43. (B,D) Whole-mount dorsal view; (C,E) transverse sections at the eye level. (F-I) In situ hybridisation analysis of Emx2 at stage 35 after injection of control (F) or Wise (G-I) morpholino oligonucleotides into a dorsal-animal blastomere at four- to eight-cell stages. (F,G) Dorsal view, anterior towards the top. (H,I) Lateral view. Wise morpholino oligonucleotides injection causes loss of olfactory placode. Arrowheads in G and H indicate unaffected placode on the uninjected side. (J,K) Injection of Wise antisense morpholino oligonucleotides causes thick surface ectoderm (* in K) in comparison with the un-injected side or control embryo (J). (L) The eye defect caused by antisense morpholino oligos can be partially rescued by co-injection of chick Wise RNA. |
