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Fig. S5 arnt2 and sim1 morpholinos effectively block correct splicing of pre-mRNA. (A) Schematic figure showing the first three exons and introns of the arnt2 gene and the splice morpholino targeting the exon2-intron2 boundary used in this study. A cryptic splice site in exon2 (depicted by an asterisk) causes deletion of 19 basepairs of the 3′ end of exon2, leading to a frameshift and a truncated Arnt2 protein. (B) Schematic figure showing the first three exons and introns of the sim1a gene and the splice morpholino targeting the exon2-intron2 boundary of sim1a. The morpholino leads to a complete loss of exon2 (83 basepairs). (C) RT-PCR from uninjected embryos (lane 1) and from embryos injected with 2.8 ng (lane 2), 3.5 ng (lane 3) or 4.5 ng (lane 4) arnt2 splice morpholino showing dose-dependent downregulation of wild-type splice pattern arnt2 transcript in injected embryos. The amount of actin transcript is unchanged. (D) RT-PCR from uninjected embryos (lane 1) and from embryos injected with 1.1 ng (lane 2) or 2.2 ng (lane 3) sim1a splice morpholino showing dose-dependent downregulation of wild-type splice pattern sim1a transcript in injected embryos. The amount of actin transcript is unchanged.