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Fig. 8. Analysis of malleal/incal joint formation in the murine middle ear. (A-C) ³⁵S radioactive in situ for Nkx3.1. (D) Section stained with Alcian Blue and chlorantine Fast Red. (E,I) ³⁵S radioactive in situ for Bapx1. (F) ³⁵S radioactive in situ for Eya1. (G,J) ³⁵S radioactive in situ for Gdf5. (H,K) ³⁵S radioactive in situ for Gdf6. (A-C,G-K) Transverse sections E14.5. (D-F) Sagittal sections E14.5. (A-C,J,K) Bapx1^-/- embryos. (A-C) Contrary to expectations, Bapx1^-/- mice display no malleal/incal joint phenotype raising the possibility of functional compensation by a related gene. ³⁵S radioactive in situ for Nkx3.1 reveals that expression is unaffected in the Bapx1 mutant head (A), with no upregulation seen in the middle ear (C). Expression of Eya1, a gene that, when inactivated, gives rise to malleal/incal joint defects, is expressed around the developing malleus and incus (F) but, in contrast to Bapx1 (E), is excluded from the region between the two ossicles. (G-I) Expression of Gdf5 and Gdf6 overlap with that of Bapx1 in the incudo-malleal joint but expression of both genes is unaffected in the Bapx1 mutant (J,K).

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