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Fig. S1 Efficacy controls of used ATG- and splice grhl1 morpholinos. (A-C) Live embryos at late blastula stages that were injected with constructs indicated at the top for genes indicated at the left side. These images show that the three ATG-MOs targeted specifically and efficiently the sequence they were designed against. (D) Graphical illustration of the grhl1-splice MO targeting site and the primer binding sites used to control the efficacy. (E) The grhl1-splice MO interfered with splicing of intron3-4, indicated by the RT-PCR amplification of an exon3-intron3/4 fragment from morphant (MO), but not from uninjected control WT embryos (WT). In reverse, a combination of an exon3 and an exon4 primer only gave an RT-PCR amplification product from WT, but not from morphant embryos (data not shown). Intron3/4 contains an in frame stop codon after the third exon (highlighted in red in D), which will yield a C-terminally truncated Grhl1 protein.