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Fig. S3 Phenotypic analysis of an S1PR2-TALEN-injected embryo. (A) An uninjected embryo (28 hours post fertilization (hpf)): Tg(cmlc2:mRFP). All observed embryos had one heart (n=64 embryos), as visualised by mRFP expression driven by a cardiac-specific promoter, cmlc2 (cardiac myosin light chain 2). No cardia bifida (two-heart phenotype) was observed. (B) An S1PR2- TALEN-injected embryo (28 hpf). Half of the embryos (34/68), exhibited cardia bifida following the injection of S1PR2-TALEN mRNA (400 pg each) into the blastomere of 1- to 2-cell stage embryos. (C) An S1PR2-MO-injected embryo (28 hpf). Nearly half of the embryos (36/66) exhibited cardia bifida following S1PR2-MO (10 ng) injection, as described previously (Kawahara et al., 2009). All images are ventral views, with the anterior at the top. Scale bars: 200 μm.