Fig. S2

Fig. S2 Morpholino-mediated knockdown of popdc2 affects muscle and heart development. (A) Genomic organization of the popdc2 gene in zebrafish. The gene consists of four exons of which exon 1–3 are coding (black box), whereas exon 4 is non-coding. The morpholino-modified oligonucleotides were directed against the splice donor (MO2) and acceptor sequences (MO1) of intron 1, respectively. Arrows depict the location of the two primer sets to amplify exon (E) and intron (I) sequences, which were utilized in the RT-PCR analysis. (B–D) RT-PCR amplification of total RNA isolated of (B) MO1-popdc2 and control morpholino-injected embryos, (CTR) respectively at 4 dpf, (C) MO2-popdc2 and control morpholino-injected embryos, respectively, at 3 dpf, and (D) MO1/MO2-popdc2 and control morpholino-injected embryos, respectively, at 2 dpf. In each case expression analysis of β-actin served as a control. The morphants displayed a significant decrease of popdc2 transcripts, and an increase in aberrantly spliced transcripts as revealed by the presence of intron 1. (E–H) Lateral view of (E) control (CTR), (F) MO1-popdc2 morphants, (G) MO2-popdc2 morphants, and (H) MO1/MO2-popdc2 morphants at 24 hpf (left) and 48 hpf (right), respectively. All morphants develop similar morphological phenotypes displaying aberrant tail morphology (arrows in F, G and H) and signs of embryonic heart failure as evidenced by blood retention and pericardial edema (arrowheads in F,G,H).