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Fig. 3 Maternal and zygotic contribution of gsc in the headless phenotype. (A) Correct and aberrant splicing of zygotic gsc pre-mRNA in the presence of morpholinos directed against the donor, Mo-gsc (D), and the acceptor, Mo-gsc (A), of splice of the first intron of the gsc gene. Exons are boxed, homeobox is in grey. P1 and P2, forward and reverse primers used for RT-PCR analysis of injected embryos. (B) RT-PCR analysis of progressive inhibition of the splicing of gsc first intron in embryos injected with a range of Mo-gsc (D) and Mo-gsc (A). The numbers on the left indicate the size of the RT-PCR products representing the aberrantly (1.1 kb) and correctly (0.7 kb) spliced RNAs. Lane 1, non-injected wild-type embryos; lanes 2–5, embryos injected with 0.6 pg, 1.2 pg, 2.5 pg and 5 pg of each morpholino, respectively; lane 6, reverse transcriptase free negative control. (C) Frequency of class I and class II phenotypes analyzed at 24 hpf in Mo-gsc (D) plus Mo-gsc (A) injected embryos (5 pg each Mo) and in embryos with an additional injection of Mo-foxA3 (10 pg). (D) Nucleic and amino acid sequence of gsc fragment generated from the aberrant splicing. Intronic region is shown in red and italic. Star indicates the stop codon.
Reprinted from Developmental Biology, 290(1), Seiliez, I., Thisse, B., and Thisse, C., FoxA3 and goosecoid promote anterior neural fate through inhibition of Wnt8a activity before the onset of gastrulation, 152-163, Copyright (2006) with permission from Elsevier. Full text @ Dev. Biol.