Rspo1 is required for normal vegfaa expression. Wild-type (A,D,G,J,M,P) or Rspo1 MO injected animals (B,E,H,K,N,Q) were examined by WISH for expression of vegfaa (A,B), shha (D,E), the Shh transcriptional target ptch2 (G,H), tgfb1a (J,K), tgfb1b (M,N), or tgfbr2 (P,Q) at the time points indicated. vegfaa, which is expressed in the medial somite of uninjected embryos, is diminished in Rspo1 MO-injected animals (A,B). shha in the hypochord and its target, ptch2, in the somite are unaffected (D,E,G,H). Tgfb1a and tgfb1b are also suppressed in Rspo1 knockdown embryos (J,K,M,N). Red arrowhead in J indicates dorsal aorta. Tgfbr2 in the somite is unaffected (P,Q). Numbers of embryos displaying the depicted phenotype are indicated. (C,F,I,L,O,R) Histograms illustrate percentages of phenotypes. Lateral views of the mid trunk at 160× magnification. (S) Diminished vegfaa expression was confirmed by qRT-PCR normalized to expression of ef1a at 20 hpf, 22 hpf, and 24 hpf (mean±s.e.m.). *P≤0.0008, determined by one-way ANOVA.
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