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Fig. 6. Misexpression of SOX9 in the gizzard mesoderm specifies the gizzard epithelium into pyloric epithelium. Histological sections of control E9 stomach (A,B), SOX9- (C) and SOX9 ${Delta}$ Cter- (D) misexpressing E9 stomachs. Immunohistochemistry analyses with ${alpha}$ 3C2 antibodies confirmed that the infection was restricted to the mesoderm of the stomach (inset in C and D) and clearly excluded from the endoderm. Control gizzard epithelium cells present keratin long cilia (A). Control pyloric sphincter epithelium cells are characterized by bulbous cilia (B). Upon mesodermal SOX9 misexpression, the gizzard epithelial cells present bleb-like cilia (C), while after mesodermal SOX9 ${Delta}$ Cter misexpression, the pyloric epithelial cells present keratin long like cilia (D). (E-G) Analyses of Pdx1 expression by in situ hybridization on section of GFP- (E), SOX9- (F) and SOX9 ${Delta}$ Cter- (G) misexpressing stomachs using antisense Pdx1 riboprobe. There is normal expression of Pdx1 in the pyloric endoderm. Ectopic Pdx1 expression is detected in SOX9-misexpressing gizzard epithelium (red arrowheads, F). In SOX9 ${Delta}$ Cter-misexpressing stomach, no Pdx1 expression is detected in the gizzard epithelium and downregulation of Pdx1 is observed in the pyloric epithelium (G). Note the expression of Pdx1 in the pancreas in these SOX9 ${Delta}$ Cter-misexpressing stomach (inset, G). duo, duodenum; e, endoderm; gizz, gizzard; m, mesoderm; panc, pancreas; pyl, pyloric structure.

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