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Na,K-ATPase α2 and Ncx4a in DFCs/KV are required for LR patterning. (A) Morpholino knockdown of genes encoding Na,K-ATPase α2 or Ncx4a in DFCs induces LR patterning defects in zebrafish. Bar chart shows the percentage of embryos injected with α2-MO or Ncx4a-MO developing cardiac looping defects. (B,C) Fluorescent signals of the α2-MO accumulated in DFCs in embryos injected with the MO at the 128-cell stage (B), but were largely excluded from DFCs when the injection was done at the dome stage (C). Arrows point to DFCs. (D,E) The expression pattern of ntl in α2 morphants (E) is similar to that observed in CTL-MO-injected embryos (D). (F,G) The morphology of KV (arrowheads) appears normal in control (F) and α2-MO-injected (G) embryos. (H,I) Anti-acetylated tubulin staining shows that cilia are present in KV of control (H) and α2-MO-injected (I) embryos.
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