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Fig. 5. An SBMO prevents mRNA maturation and causes developmental abnormalities in embryos. (A) Schematic showing oligo binding site and RT-PCR demonstrating inhibition of mRNA maturation by the SBMO. Both pseudoalleles are affected. Genomic DNA template was used as a positive control for unspliced product. (B) An alternative morpholino, targeting the splice donor site, does not efficiently inhibit mRNA maturation. (C) RTPCR analysis of XBra and FGF8 expression in animal caps depleted of FoxI1e and treated with 1 g/ml Activin A. SBMO-injected animal caps showed levels of mesoderm induction similar to uninjected caps. (D,E) Embryos injected animally at the two cell stage with a dose range of SBMO. There were dose-dependent abnormalities in development, shown at the neurula stage (stage 19, D) and in tailbud stage embryos (stage 27, E). (F) Epidermal cytokeratin, slug, sox-2, FoxG1 (BF-1) and Xlhbox6 are all reduced at stage 23. E. keratin, slug and sox- 2 are partially rescued by 50 pg FoxI1e mRNA. (G) Xbra, Xwnt-8 and chordin are unaffected by SBMO injection at stage 11.5. Image published in: Mir A et al. (2007) Copyright © 2007. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Experiment + Assay Source Phenotypes and Disease Xla Wt + Activin + animal cap explant + NF11 (RT-PCR) fig.5.c Expression Phenotype increased amount fgf8.L expression in animal cap increased amount tbxt.L expression in animal cap Xla Wt + foxi1 MO + Activin + animal cap explant + NF11 (RT-PCR) fig.5.c Expression Phenotype increased amount fgf8.L expression in animal cap Xla Wt + foxi1 MO + NF19 (morphology) fig.5.d Anatomical Phenotype abnormal development of cement gland abnormal development of neural tube Xla Wt + foxi1 MO + NF27 (morphology) fig.5.e Anatomical Phenotype abnormal axis elongation abnormally incomplete structure of epidermis decreased length of anterior-posterior axis decreased size of the head Xla Wt + foxi1 + foxi1 MO + NF23 (in situ hybridization) fig.5.f Expression Phenotype decreased amount krt12.4.L expression in embryo decreased amount snai2.L expression in embryo decreased amount sox2.S expression in embryo Xla Wt + foxi1 MO + NF23 (in situ hybridization) fig.5.f Expression Phenotype decreased amount foxg1.L expression in embryo decreased amount hoxb9.L expression in embryo decreased amount krt12.4.L expression in embryo decreased amount snai2.L expression in embryo decreased amount sox2.S expression in embryo Larger Image Printer Friendly View

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