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Fig. 1. Developmental failure of the frontal bone primordium in Tgfbr2^fl/fl;Wnt1-Cre mice. (A,B) E16.5 embryo skeletal preparation reveals severe defects of frontal (fr), parietal (pr) and interparietal (ip) bone in the Tgfbr2^fl/fl;Wnt1-Cre mutant. Only the orbital aspect of the frontal bone was developed in the Tgfbr2^fl/fl;Wnt1-Cre mutant. (C,D) At E12.5, the CNC-derived mesenchyme begins to form a condensation (arrow) that serves as the template for frontal bone development in both the wild type and the Tgfbr2^fl/fl;Wnt1-Cre mutant. (E,F) A well-defined, CNC-derived frontal bone primordium is clearly visible (arrow) in the wild-type sample at E13.5. In the Tgfbr2^fl/fl;Wnt1-Cre mutant, the development of the frontal bone primordium is retarded when compared with the wild-type sample (arrow). (G,H) At E14.5, a well-developed bone matrix is visible within the frontal bone primordium of the wild-type sample. In the Tgfbr2^fl/fl;Wnt1-Cre mutant, however, bone matrix fails to form within the frontal primordium (arrow), while the orbital aspect of frontal bone is present (arrowhead). (I,J) At E16.5, frontal bone formation is evident in the wild-type sample (arrow). No bone formation is detected in the calvarial aspect (arrow) of the frontal primordium in the Tgfbr2^fl/fl;Wnt1-Cre mutant. (K,L) At birth (NB), the frontal bone is well developed to form the roof of bony orbit (arrowhead) and covers the side of skull (arrow). In the Tgfbr2^fl/fl;Wnt1-Cre mutant, the frontal bone development is retarded with a rudiment of orbit surface region (arrowhead), whereas there is no development of the calvarial aspect of frontal bone (arrow). Scale bars: 1 mm in A,B; 200 µm in C-L.

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