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Fig. 5. Shh signaling is restored in the limb bud of N-Shh rescued Disp1-null mutants. Whole-mount in situ hybridizations with antisense riboprobes for Shh (A-C) and Ptch1 (D-F) in limb buds of wild-type (A,D), N-Shh^C/Shhⁿ; Sox2Cre (B,E) and N-Shh^C/Shh^n; Sox2Cre; Disp1^C829F/C829F (C,F) embryos at 10.5 dpc as indicated. Shh gene expression is restricted to the posterior mesenchyme in the wild type (A). N-Shhp from this domain diffuses anteriorly, which then leads to a graded Ptch1 expression across the entire posterior half of the limb field (D). The expression of N-Shh is induced in the posterior mesenchyme in N-Shh mutant (B). Ptch1 is reduced and restricted to the posterior mesenchyme (E). High levels of Ptch1 expression are restricted to Shh-producing cells and to cells immediately anterior to this domain. No gradient of expression is apparent across the AP axis. Similar levels of Shh and Ptch1 expression are observed in limb buds of N-Shh rescued Disp1 mutant embryos (C,F).

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