Fig. 2.

Expression of Neo1 and limb patterning genes in control and Neo1^Gt/Gt mice. A: Expression of the Neo1-lacZ reporter in E10.5 and E11.5 hindlimb buds. B: Western blot analysis of neogenin expression in E11.5 wild-type and Neo1^Gt/Gt limb buds and in mouse embryo fibroblasts (MEFs). Blots were probed with neogenin and, as a loading control, pan-cadherin antibodies. Numbers below lanes represent the relative amounts of neogenin produced by limb buds of the indicated genotype, normalized to cadherin expression. A similar analysis was performed with MEFs, which were normalized independently from limb buds. C: Whole mount in situ hybridization of Shh (n=8), Gli1 (n=8), Ptch1 (n=10), Bmp4 (n=10), Msx1 (n=12), Msx2 (n=12) and Alx4 (n=10) expression in control (Neo1^+/+ or Neo1^+/Gt) and Neo1^Gt/Gt right hindlimb buds at E11.5. Red arrowheads indicate ectopic anterior expression of Gli1 and Ptch1.