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Fig. 5. Grem1, but not SHH, rescues Fgf8 and Bmp2 expression in the AER of Gre{Delta}ORF/{Delta}ORF limb buds. All grafted limb buds are forelimb buds (E10.5) of Grem1 mutant embryos. Limb buds either received Shh (red arrow) or Grem1 (green arrow)-expressing cell aggregates and were cultured for 15 hours prior to analysis. White arrowheads indicate the endogenous expression domains; blue arrowheads and asterisks indicate the induced expression. (A) Bmp2 expression in a non-grafted control limb bud of a mutant embryo. (B) Posterior grafts of Shh-expressing cell aggregates fail to rescue Bmp2 expression. (C,D) Posterior grafts of Grem1-expressing cells induce Bmp2 expression in the AER (D), while no Bmp2 transcripts are detected in the AER of non-grafted mutant limb buds (C). Note also the enhancement of mesenchymal Bmp2 expression (D). (E,F) Posterior grafts of Shh-expressing cells do not rescue Fgf8 expression in the AER (F) in comparison with a non-grafted mutant limb buds (E). (G,H) Posterior grafts of Grem1-expressing cells induce upregulation of Fgf8 expression in the AER (H) in comparison with endogenous Fgf8 expression in non-grafted mutant limb buds (G). (A-D) Dorsal views with posterior towards the bottom and distal towards the right; (E-H) posterior is towards the bottom and dorsal towards the left.