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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 3.

Figure 3

EAD ectoderm undergoes convergent extension (CE) as the cranial neural crest (NC) approaches the midline and EAD CE fails to occur in sox9 LOF embryos. (A) Experimental schematic. (B-D’’) Coronal sections with mGFP-labeled NC (green) and β-catenin (red) immunolabeling from late neurula (stage 24) to late tailbud (stage 28) (2 independent experiments, st. 24, n=7; st. 26, n=7; st. 28 n=4). Midline region (bracket) with bright β-catenin labeling is EAD ectoderm. Bracket: region of 10x image (B-D) enlarged in 40x view (B’-D’) and 63x view (B’’-D’’). cg, cement gland. (E-F) Frontal view of control and sox9 LOF embryos at swimming tadpole (stage 40) assayed in 2 experiments (control MO (E) n=24; sox9 MO (F) n=30.) Dots surround open mouth. Bracket: unopened mouth. Scale bar: 200 m. (G-H’) Coronal sections assayed in 4 independent experiments (n=23) with β-catenin immunolabeling and Hoersch nuclear labeling. Midline region with bright β-catenin labeling is EAD ectoderm. Bracket: region of 10x image (G-H) enlarged in 25x view (G’-H’). cg, cement gland. (I) Graph depicting percent of embryos displaying mouth, face, nostrils and pigment formation phenotypes at st. 40 in control and sox9 LOF embryos. P values: Fisher's exact probability test. (J) Quantification of height over width of EAD (see Methods). P values: unpaired, two-tailed T test. Error bar: standard deviation. Unless otherwise specified, Scale bar (10x): 170μm. Scale bar (25x): 68μm. Scale bar (40x): 43μm. Scale bar (63x): 27μm.