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Fig. S4
Interactions between Wnt4a, Wnt11r, and Fgf8a during pouch formation
(A-F) Alcama immunohistochemistry at 34 hpf shows a series of five pouches. Compound wnt11r+/-; fgf8a-/- but not wnt4a-/-; fgf8a-/- mutants show enhancement of pouch defects compared to fgf8a-/- embryos alone. Sensory ganglia are indicated with red asterisks. (G-L) Facial cartilages stained with Alcian. Compound wnt11r+/-; fgf8a-/- mutants but not wnt4a-/-; fgf8a-/- mutants show enhancement of ceratobranchial cartilage defects compared to fgf8a-/- mutants alone. (M) Quantification of pouch and ceratobranchial cartilage (CB) defects. Number of pouches and CBs examined for each genotype: wild type (106, 102), wnt11r+/- (51, 47), wnt4a-/- (45, 144), fgf8a-/- (68, 98), wnt11r+/-; fgf8a-/- (38, 80), and wnt4a-/-; fgf8a-/- (20, 30). Data represent mean ± SEM. ***, p<0.001. n.s., not significant. The data for wild type and fgf8a-/- are repeated from Figure 4I. (N-Q) Fluorescent in situ hybridization for fgf8a or wnt11r mRNA (green) at 30 hpf. GFP immunohistochemistry labels the nkx2.5:GFP+ mesoderm in red. fgf8a expression is unaffected in 10/13 and slightly reduced in 3/13 wnt11r mutants. wnt4a expression is unaffected in 15/19 and slightly reduced in 4/19 fgf8a mutants. Scale bar represents 40 μM