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Fig. 2. Early hindbrain expression of Hoxb4 and Rarb requires Raldh2 activity. The effects of loss of Raldh2 activity on the early neural expression of Hoxb4 mRNA (A,B), Hoxb4 protein (C,D) and Rarb gene (E,F) are shown. (A,B) Dorsal views of E8.5 embryos showing the Hoxb4 mRNA distribution in Raldh2^+/+ (A) and Raldh2^-/- (B) genetic backgrounds. Loss of Raldh2 function is associated with absence of Hoxb4 mRNA within the hindbrain but expression within the developing spinal cord (sc) remains unaffected. (C,D) Dorsal views of E8.5 embryos showing Hoxb4 protein expression in Raldh2^+/- (C) and Raldh2^-/- (D) embryos. Removal of Raldh2 activity leads to loss of most Hoxb4 protein within both the developing hindbrain (hb) and spinal cord (sc). However, weak expression is still observed within a few cells at caudal hindbrain and anterior spinal levels. (E,F) Rarb mRNA induction within the hindbrain requires Raldh2 activity. Lateral views of E8.5 embryos of the genotypes Raldh2^+/- (E) or Raldh2^-/- (F). Loss of Raldh2 activity is associated with the absence of all detectable Rarb expression within the developing hindbrain and spinal cord. Os and Ov indicate the otic sulcus and otic vesicle, lying adjacent to r5 and r6 in this and subsequent figures, and the approximate position of the paraxial mesodermal limit of Hoxb4 expression at the somite 6/7 border is also shown (horizontal line in A-D).

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