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Fig. 3. Induction of cardiac precursor gene expression is unaffected in morphant Xenopus neurulae. (A) Nkx2.5 was expressed normally at stage 16/17 when GATA4, 5 or 6 were depleted individually or in combination. (B) Cryostat sections confirmed that expression of Nkx2.5 in the cardiac mesoderm (delineated by red dashed lines) was not affected. (C) Expression of Nkx2.3, which unlike Nkx2.5 is restricted to the cardiac mesoderm (D) also remained unchanged in the triple morphants, as does eHAND (E). (Red dashed lines mark the cardiac precursors, remaining stain reflects expression in the blood island mesoderm). |
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Fig. 1. Cardia bifida is evident in GATA4, 5 and 6 morphants, but cardiac gene expression is only affected in GATA6 depleted Xenopus embryos. (A) Cardiac tissue stained for MLC fails to migrate to the midline in embryos injected singly with GATA4, 5 or 6 morpholinos. (B, C) Expression levels of MLC, Nkx2.5, Tbx5 and CA remain unchanged in GATA4 and 5 morphants compared with control uninjected embryos at stage 28. (D) Expression levels of MLC, Nkx2.5, Tbx5 and CA are substantially decreased in GATA6 morphants at stage 28. |
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Fig. 2. Functional redundancy between GATA4, 5 and 6. (A) Xenopus embryos were injected singly and in combination with GATA4, 5 and/or 6 morpholinos, harvested at stage 28 and analysed by whole mount in situ hybridisation for MLC and Nkx2.5 expression. Morphant embryos were classified into four classes: wild type (type +, light blue bar), mild down regulation of MLC or Nkx2.5 (type −, mid blue bar), strong down regulation (type −−, purple bar) and no expression (type −−−, dark blue bar). (B, C) Graphical representations of the proportion of embryos in each class. |
