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Fig. S2 Splicing of irxl1 is disrupted in MOII morphants. A: Locations of the irxl1 morpholino oligos (MO). MOI is complementary to 25 bp spanning the translation initiation site of irxl1 mRNA (-17 to +8), MOII is complementary to 25 bp spanning the boundary of intron 3 and exon 4. B: Reverse transcriptase-polymerase chain reaction analysis of RNA isolated from wild-type and MOII-injected embryos at 25 hpf using primers a, b, c as indicated in (A). Primers a+b will amplify the correctly-spliced transcript (381 bp) whereas primers a+c will amplify a transcript with intron 3 unspliced (predicted size 344 bp). The correctly-spliced product (381 bp) is significantly reduced in MOII morphant, while the unspliced product (344 bp) is present in MOII morphant but not wild-type embryos, indicating that splicing of irxl1 is disrupted in MOII morphants.