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Fig. 6. Fgfr2, Dlx5, and Twist1 are downstream targets of TGFß signaling in regulating frontal bone development. (A) At E13.5, the expression of Fgfr2 is detected in the frontal primordium (arrow) of the wild type. (B) The expression of Fgfr2 is reduced in the frontal primordium (arrow) of the Tgfbr2fl/fl;Wnt1-Cre mutant. (C) At E14.5, frontal bone matrix begins to form within the frontal primordium of the wild-type sample. The expression of Fgfr2 is detected throughout osteogenic progenitor cells in the frontal bone primordium (arrow) and the orbital surface region of the frontal bone (arrowhead). (D) In the Tgfbr2fl/fl;Wnt1-Cre mutant, there is virtually no Fgfr2 expression in the calvarial aspect of the frontal primordium (arrow), while residual Fgfr2 expression is detected within the orbital aspect of the frontal bone in the Tgfbr2fl/fl;Wnt1-Cre mutant sample (arrowhead). (E) At E13.5, Dlx5 is expressed in the CNC-derived frontal bone primordium (arrow). (F) Comparable Dlx5 expression is present in the frontal primordium in the Tgfbr2fl/fl;Wnt1-Cre mutant (arrow). (G) The frontal bone matrix begins to form at E14.5. Dlx5 is expressed in both the calvarial (arrow) and the orbital (arrowhead) aspects of the frontal bone primordium. (H) The Tgfbr2fl/fl;Wnt1-Cre mutant fails to express Dlx5 in the calvarial aspect of the frontal bone primordium, whereas expression on the orbital aspect (arrowhead) is normal. (I) No Twist1 expression is detected in the frontal primordium in the wild-type sample. (J) Elevated Twist1 expression is detected in the frontal bone primordium (arrow), whereas there is no Twist1 expression in the orbit region of the frontal bone anlagen (arrowhead) in the Tgfbr2fl/fl;Wnt1-Cre mutant. Scale bar: 200 µm.