Figure 2.

actn2 MO effectively knocks down the actn2 gene and induces a heart-specific phenotype. A) Schematic of the gene structure of actn2 and the exons that are targeted by the three splice donor MOs E1 MO, E6 MO, and E7 MO. actn2 MO (bold) refers to E7 MO. B) Injection of actn2 MO induces an exon-skipping event, as revealed by DNA gel electrophoresis after RT-PCR using a primer pair spanning exons 6 and 8. The 246-bp band detected in the WT fish is shifted to 165 bp in the actn2 MO fish, indicating the deletion of exon 7. C) Western blot analysis indicates a strong reduction in Actn2 protein in hearts dissected from embryos that were injected with either E7 MO or E6 MO. D) Lateral views of zebrafish embryos show that embryos injected with actn2 MO exhibit a cardiac-specific phenotype. Pericardial edema (arrowhead) can be rescued by Tg(ttna:actn2-egfp).