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Fig. 5. Molecular defects in osteoblast development in the DKO embryo. (A1-E3) In situ hybridization using ³⁵S-labeled riboprobes against Col10a1 (A1-A3), Runx2 (B1-B3), Ap (C1-C3), Osx (D1-D3) and Bsp (E1-E3), on longitudinal sections through the humerus from E15 embryos of wild type (A1-E1), Ihh^-/- (A2-E2) or DKO (A3-E3). Shown are the distal regions of the humerus for the wild type (A1-E1) and the DKO (A3-E3) embryo, but the full-length for the Ihh^-/- embryo (A2-E2). Hybridization signals are in red. Red and green arrows indicate signals in the perichondrium; red and green asterisks indicate signals in chondrocytes (see text). e, epiphysis; d, diaphysis; t, tendon. (F1,F2,G1,G2) In situ hybridization using ³⁵S-labeled riboprobes against Tcf1 (F1,F2) and Dkk1 (G1,G2) on longitudinal sections through the humerus from E14.5 wild type (F1-G1) and DKO (F2,G2) embryos. Shown are the distal regions of the humerus. Red arrows indicate the perichondrium. (H1,H2) In situ hybridization using ³⁵S-labeled riboprobes against Tcf1 on longitudinal sections through the humerus from E18.5 embryos of wild type (H1) and DKO (H2). Red arrows indicate either signals in the perichondrium (H1) or the corresponding regions missing the signals (H2). Red asterisk (H2) indicates the ectopic bone. H, hypertrophic zone; PS, primary spongiosa.

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