Wnt/β-catenin signaling is essential for the LR patterning function of KV. (A-F) charon expression was suppressed in wnt3 (D), wnt8 (E), and wnt3/wnt8 double (F) morphants, but not in embryos overexpressing wnt3 (B) or wnt8 (C) RNA, as compared with control embryos (A). Shown are dorsal views of tail bud regions at the 12-somite stage. (G-N) Cilia development in KV was regulated by Wnt signaling, as shown by anti-acetylated tubulin antibody staining of 10-somite staged embryos. Compared with controls (G; n=15), shorter KV cilia were observed in wnt3 morphants (J; n=15), wnt8 morphants (K; n=12), and wnt3/wnt8 double morphants (L; n=15), but not in embryos injected with wnt3 RNA (H; n=12) or wnt8 RNA (I; n=12), as summarized in M. Cilia numbers were increased in embryos overexpressing wnt3 (H) or wnt8 (I) RNA but remained the same in wnt3 (J), wnt8 (K) and wnt3/wnt8 double (L) morphants, as summarized in N. Data shown are means±s.d. *P<0.001, Student's t-test.