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Fig. 7. Wnt is necessary for inner ear formation before 9ss of development. (A) Soho1 expression is reduced as a result of Dkk1 electroporation (n=12/18). (B) Section shows a reduction of Soho1 and thickened otic ectoderm on one side of the embryo. (C) mCherry tracer expression, as revealed by dsRed immunoreactivity, is unilateral and is seen throughout the ectoderm. (D) Nkx5.1 is also reduced by Dkk1 overexpression. (E) A lateral view of the embryo shown in D, showing widespread mCherry expression throughout the ectoderm. For clarity, the outline of the embryo has been traced. The heart tube (h) can be visualised. (F) Despite unilateral Dkk1 electropoartion, Pax2 expression in the OEPD is initially normal at 7ss (n=8/9 showing normal expression). (G) Sections show normal ectodermal Pax2 expression. (H) Electroporated cells are revealed using dsRed immunostaining. (I) At 13ss, Pax2 is downregulated in a medial portion of the Pax2 expression domain (n=4/5). Line marks the axial level of the section. (J) Section of the embryo shown in I, showing a reduction of Pax2 medially (marked by the bar). (K) Section of the embryo shown in I, showing that electroporation of Dkk1 targets a large extent of the ectoderm, as shown by dsRed immunoreactivity. (L) Foxi2 expression encroaches into the otic territory (n=5/8). (M) Section through embryo shown in L. The region of Foxi2 otic exclusion is reduced by 50%. (N) dsRed immunoreactivity shows the extent of electroporation. (O,P) Epibranchial neurogenesis as marked by Phox2b is not obviously affected as a result of Dkk1 electroporation when control (O) and electroporated (P) sides are compared.

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