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. Author manuscript; available in PMC: 2017 Aug 2.
Published in final edited form as: Cell Rep. 2016 Jul 14;16(5):1445–1455. doi: 10.1016/j.celrep.2016.06.073

Figure 4.

Figure 4

Fzl7 is locally required in the EAD ectoderm for convergent extension. Local requirement of Dsh, fzl7, and wnt11 expression tested with an EAD transplant technique. (A) Experimental design: donor LOF tissue was transplanted to uninjected sibling recipients. (B-C’) EAD transplant outcome from control or Dep+ RNA donor tissue assayed in 3 experiments. ((B, B’) control RNA n=23; (C, C’) Dep+ RNA n=22.) (B’-C’) Overlay of (B-C) with GFP fluorescence indicating location of donor transplant in recipient. Dots surround open mouths. Bracket: unopened mouth. Frontal view. cg, cement gland. Scale bar: 200 m. (D-E’) Coronal sections of EAD transplants with control or Dep+ donor tissue assayed in 3 independent experiments ((D, D’) control RNA n=10; (E, E’) Dep+ RNA n=14) with β-catenin immunolabeling. Midline region with bright β-catenin labeling is EAD ectoderm. Bracket: region of 10x image (D-E) enlarged in 25x view (D’-E’). (F) Quantification of normal or abnormal structure development depending on background of facial tissue. P values: Fisher's exact probability test. (G) Quantification of height over width of EAD (see Methods). P value: unpaired, two-tailed T test. Error bar: standard deviation. (H-J) EAD transplant outcome from control, fzl7 or wnt11 LOF donor tissue assayed in 4 independent experiments. ((H, H’) control MO n=27; (I, I’) fzl7 MO n=30; (J, J’) wnt11 MO n=30.) (H’-J’) Overlay of (H-J) with GFP fluorescence indicating location of donor transplant in recipient. Dots surround open mouths. Bracket: unopened mouth. Frontal view. Scale bar: 200μm. (K-M’) Coronal sections of EAD transplants with control, fzl7 or wnt11 donor tissue assayed in 4 independent experiments with β-catenin immunolabeling ((K, K’) control MO n=19; (L, L’) fzl7 MO n=17; (M, M’) wnt11 MO n=14). Midline region with bright β-catenin labeling is EAD ectoderm. Bracket: region of 10x image (K-M) enlarged in 25x view (K’-M’). (N) Quantification of normal or abnormal structure development depending on LOF background of facial tissue. P values: Fisher's exact probability test. (O) Quantification of height over width of EAD midline tissue in transplants. P values: unpaired, two-tailed T test. (P) Schematic of model. NC releases Wnt11 which acts on Fzl7 receptors expressed on midline EAD cells. Unless otherwise specified, Scale bar (10x): 170μm. Scale bar (25x): 68μm.