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Fig. 2. Expression of the Nkx2.9-lacZ reporter gene in heterozygous (A-E) and homozygous (F) Nkx2.9 mutants. lacZ staining of whole-mount embryos at E8.5 (A), E9.5 (B), E11.5 (C) and E12.5 (D). Insets in A and B show whole-mount in situ hybridization with Nkx2.9-specific probe. Note the slightly delayed onset of lacZ activity compared with endogenous Nkx2.9 transcription in the E8.5 embryo (A) but a comparable expression in the E9.5 embryo (B). lacZ activity continues to be present in Nkx2.9 domains of embryos at E11.5 (C) and E12.5 (D). Comparison of lacZ staining in E10.5 heterozygous (E) and homozygous (F) mutants reveals identical activity patterns except for the enlarged domain in hindbrain of homozygous Nkx2.9 mutants at the level of rhombomeres 3 and 4 (arrows in E,F). Inset in E,F shows lacZ staining on transverse sections of spinal cord at the forelimb level.