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Figure 4


Fig. 4. Expression of otic expressed genes in the ectopic vesicles at the level of the hindbrain. (A) Spalt4, (B) Sox10, (C) Notch1, (D) Lunatic fringe, (E) EphA4, (F) Tbx1, (G) Dlx5. (H) Six1 mRNA was not detected in an ectopic vesicle by in situ hybridization (arrow), though surrounding cells were positive. (I) GFP. (J) An antibody to Spalt4. The same section as in I. (K) GFP. (L) Same section as in K showing Pax2 expression. (M) GFP. (N) The same section as in M showing Dlx3 expression. (O) An ectopic vesicle in an embryo 72 hours post electroporation, with the Spalt4 expression construct expressing GFP (green) but there is little detectable TUJ1 staining (red). A portion of an adjacent ganglion shows strong TUJ1 staining. A-F show section in situ hybridizations. G and H are 20 µm cryostat sections of whole-mount in situ hybridizations. The embryos In I-O were paraformaldehyde fixed 48 hours (I-N) or 72 hours (O) after electroporation then sectioned at 10 µm on a cryostat and used for immunohistochemistry. Scale bars: 50 µm.





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