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Fig. S6
Testing of mych morpholino functionality. (A-H) The functionality of the mych translation-blocking morpholino (ATG-MO) was tested by injecting a fusion mRNA, where the MO target sequence was fused to the gfp ORF at the start ATG, together with different concentrations of the ATG-MO into one-cell stage embryos. The GFP signal was analyzed using fluorescence microscopy (left panel) and the normal morphology of the embryos after morpholino injection was documented using transmitted light microscopy (right panel). The translation of gfp was completely blocked by injecting as little as 1.4 ng of the ATG-MO (C). For the splice-blocking morpholino (Sp-MO) the functionality was tested by RT-PCR using a pair of primers overlapping the second intron (I), whose splicing sites are targeted by the mych-Sp-MO. In WT the 162 bp fragment reflects the proper splicing of the pre-mRNA, whereas after the injection of 4.1 ng or more of mych-Sp-MO the detected fragment contains the intron and its size increased to 396 bp (I).