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Fig. 2 A tgfbi splice morpholino efficiently blocks normal splicing of tgfbi mRNA. Normally spliced tgfbi mRNA in wild type embryos produced a single amplification product by RT-PCR (A, wt lane; B-a), whereas the same primers generated three different products from morphant embryo mRNA (A, MO lane). Sequencing of each band revealed that none derived from an mRNA encoding full-length Tgfbi protein. The top PCR product includes an insertion of intron between exon 4 and 5 with a stop codon in the intron (B-b). The second PCR product contains an insertion of a part of an intron, which causes the introduction of a stop codon in exon 5 (B-c). The third PCR product is produced by exon 4 skipping, which introduces a stop codon in exon 5 (B-d).