Fig. 2
Edn1 functions downstream of wdr68 for craniofacial development.
(A-D) Ventral views of 5dpf Alcian stained craniofacial cartilages of zebrafish raised at 32°C. A) wildtype sibling injected with EF1a mRNA. B) wdr68 mutant injected with EF1a mRNA. C) wdr68hi3812/hi3812 mutant injected with Edn1 mRNA. D) wdr68hi3812/hi3812 mutant injected with Flag-Wdr68 (FW) mRNA. E) Edn1 mRNA-injected mutants have more M cartilage elements than EF1a controls (p<0.001). (F-Q) ISH analysis of dlx6a expression in embryos raised at 32°C with red arrowhead pointing at 1st arch CNCC territory. F, H, J, L) dorsal view of prim-5 stage. G, I, K, M) lateral view of prim-5 stage. F, G) wildtype sibling injected with GFP/dsRed (G/R) plasmid mix showing normal dlx6a expression. H, I) wdr68hi3812/hi3812 mutant injected with G/R plasmid mix showing loss of dlx6a in 1st arch CNCC. J, K) wildtype sibling injected with GFP/Edn1 (G/Edn1) plasmid mix showing near-normal dlx6a expression. L, M) wdr68hi3812/hi3812 mutant injected with G/Edn1 plasmid mix showing partial rescue of dlx6a in 1st arch CNCC. N-Q) lateral view of dlx6a expression in prim-5 stage embryos raised at 28.5°C. N) heat shocked wildtype sibling control with normal dlx6a. O) heat shocked wildtype sibling injected with Et1-MO showing loss of dlx6a. P) heat shock induced Tg(hsp70l:GFP-Wdr68) sibling control with normal dlx6a. Q) heat shock induced Tg(hsp70l:GFP-Wdr68) sibling injected with Et1-MO showing loss of dlx6a.