Fig. 2

Cnpy1 function within DFCs is essential for ciliogenesis and LR patterning. (A and B) A-tubulin (green) and nucleus (red) staining in uninjected (A) or DFC^cnpy1-MO–injected (B) embryos at the six-somite stage. Vegetal pole view. (Scale bars: 20 μm.) (A2 and B2) X–Z view around the KV. Lumen formation was not completed in DFC^cnpy1-MO–injected embryos (B2). (C) Number (red) or length (blue) of KV primary cilia in uninjected (n = 10 or 49), DFC^cnpy1-MO (n = 10 or 48), yolk^control-MO (n = 11 or 77), yolk^cnpy1-MO (n = 11 or 58), DFC^{cnpy1-MO+mRFP} (n = 10 or 61), or DFC^{cnpy1-MO+Cnpy1} (n = 11 or 85) embryos. (Error bars show SEM.) Statistically significant (P < 0.05) differences could be seen in uninjected versus DFC^cnpy1-MO and DFC^{cnpy1-MO+mRFP} versus DFC^{cnpy1-MO+Cnpy1} embryos. (D and E) Representative images of mlc2a expression demonstrating normal looping (uninjected; D) or reversed looping (cnpy1-MO; E) of the heart in embryos at the high pec stage. Ventral view, anterior to the top. A, atrium; V, ventricle. (F) Percentages of normal looping, reversed looping, no looping, or cardia bifida of the heart in uninjected (n = 164), control-MO (n = 118), cnpy1-MO (n = 119), fgf8-MO (n = 65), DFC^control-MO (n = 95), DFC^cnpy1-MO (n = 146), DFC^cdh1-MO (n = 106), yolk^control-MO (n = 96), yolk^cnpy1-MO (n = 94), DFC^{cnpy1-MO+mRFP} (n = 136), and DFC^{cnpy1-MO+Cnpy1} (n = 165) embryos. Statistically significant (P < 0.05) differences could be seen in uninjected versus cnpy1-MO, DFC^control-MO versus DFC^cnpy1-MO, and DFC^{cnpy1-MO+mRFP} versus DFC^{cnpy1-MO+Cnpy1} embryos.