Fig. 6. Component and merged micrograph images for Fig. 2 K, L, and M.

Fig. 7. Lef1 null mice maintain normal morphology of their circumvallate papilla. Developing tongues from E14.5 wild-type (A) or Lef1 null (B) mice were examined by scanning electron microscopy. (Scale bar: 100 mm.)

Fig. 8. LiCl up-regulates Shh expression in organ culture. (A) Standard organ culture conditions. (B and C) Cultures with 5 mM NaCl. (D-F) Cultures with 5 mM LiCl. Note that images A and D are the same as in Fig. 4A.

Fig. 9. Inhibitors of Shh signaling increase b-gal expression in organ cultures of tongues from TOPGAL mice. (A and D) Standard organ culture conditions. (B and E) Cultures contained the anti-Shh antibody 5E1. (C and F) Cultures contained 5 mM cyclopamine.

Fig. 10. Exogenously added Shh inhibits production of endogenous Shh mRNA in organ culture. Shh transcripts in fungiform papillae of E12.5 tongues cultured for 2 days were detected by in situ hybridization of serial sections. Tongues in organ culture were treated as follows: A-C, standard conditions; D-F, contained Shh-N. Shh-positive transcripts were observed in sections from standard culture conditions (A-C, arrows) but not from cultures containing Shh-N. (Scale bar: 100 mm.)

Fig. 11. Shh inhibitors do not enhance fungiform papilla formation or up-regulate endogenous Shh in fungiform papilla of Lef1 null mice. E12.5 embryonic tongues were cultured for 2 days in standard medium (A and B) or in standard medium with anti-Shh antibody 5E1 (C and D). (A and C) Wild-type tongue. (B and D) Lef1 null tongue. The addition of 5E1 antibody increased Shh expression and fungiform papillae formation in tongues from wild-type mice but not in tongues from Lef1 null mice.