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Fig. S3 Mosaic expression of dominant-negative Rho kinase disrupts LR development. (A,B) The majority of embryos (93%, n=82) injected with 20 pg of control plasmid encoding yellow fluorescent protein (YFP) appeared normal at 2 dpf, including normal axis elongation (A). Most embryos injected with 20 pg of DNA encoding dominant-negative Rho kinase (DN-Rock) also showed normal axis elongation (75%, n=70), but some embryos developed mild (16%, n=70) or severe (9%, n=70) axial defects (B). (C,D) Asymmetric spaw expression in LPM was normal in the majority of control YFP DNA-injected embryos (C), but was altered in many DN-Rock embryos (see Fig. 4D), and was often bilateral (red arrows in D). Expression of shh in the midline was intact in uninjected embryos (n=94/94), control YFP DNA-injected embryos (n=59/59) and DN-Rock DNA-injected embryos (n=89/91). (E-H) Whereas heart looping was normal in most control embryos (E,H), DN-Rock embryos showed an increased frequency (H) of reversed (F) or no heart looping (G). Embryos with axial defects were excluded for this analysis.