Fig. S5

Fig. S5 Cardiac-specific expression of Gα₁₃a fails to rescue cardia bifida caused by global Gα₁₃ inhibition. The transgene myl7:gna13a-IRES-nlsGFP was created using the Gateway system (Kwan et al., 2007; Villefranc et al., 2007), such that Gα₁₃a is expressed specifically in the myocardial cells under control of the myl7 promoter (Huang et al., 2003). The expression of Gα₁₃a was monitored as nuclear GFP (nlsGFP) expression driven by an internal ribosomal entry site (IRES) (Kwan et al., 2007). The transgene plasmid DNA (50 pg) was co-injected with transposase RNA (30 pg) into the cytoplasm of wild-type embryos at the one-cell stage. A stable line expressing Gα₁₃a in the heart was identified. Injections of gnb13ab MOs in Tg(myl7:gna13a-IRES-nlsGFP) embryos still exhibited cardia bifida. (A) A representative epifluorescence image of 30 hpf-Tg(myl7:gna13a-IRES-nlsGFP) embryos, showing expression of nlsGFP in the heart (arrow). (B) A confocal Z-stack image showing that nlsGFP is expressed in all cardiomyocytes of Tg(myl7:gna13a-IRES-nlsGFP) embryos at 30 hpf. (C) A representative epifluorescence image of 30 hpf-Tg(myl7:gna13a-IRES-nlsGFP) embryos injected with gna13ab MOs showing cardia bifida (arrows). (D) Frequencies of cardia bifida in 30-hpf embryos. #P>0.5 versus control. Scale bar: 100 μm.