DEV02339 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Shh expression pattern in mes/r1 development. (A-C) RNA in situ hybridization. (A) At E8.5, Shh is expressed in the notochord (N) and weakly in the overlying ventral midline (VM) of the mes. fb, forebrain. (B) At E10.5, Shh is expressed in a broad ventral domain of the mes. V, ventral; D, dorsal. (C) Black arrowhead: Shh expression is maintained in the VM of the ventral mes (vmes) at E18.5. Red arrowhead: Shh expression in the Purkinje cell layer of the cerebellum (Cb). Scale bars: 25 μm in A,B; 150 μm in C.

• Supplemental Figure 2 -

  Fig. S2. Analysis of the recombination pattern in the Nestin-Cre mice using the R26R lacZ reporter allele. X-gal staining (blue) reveals the recombined areas (A) At E10.0, recombination has occurred in r1 (mainly ventral), but not in the mes. (B) By E11.5, recombination is observed throughout the mes/r1, except near the isthmus (arrowhead) and the ventral (not shown) and dorsal (arrow) midline of the mes. (C) By E12.5, recombination is almost complete in the isthmic region (arrowhead). Scale bars: 75 μm.

• Supplemental Figure 3 -

  Fig. S3. Fate-mapping of mes/r1-derived structures in Smo-En1 cko mutants. (A,B) Smo-En1 cko mutant and control embryos (En1Cre^+/-, Smo ^fl/+) on R26RlacZ background. X-gal staining (blue) demarcates the mes/r1-derived structures. (C,D) In situ hybridization for Dmbx1, which is expressed in the superficial layer of the superior colliculus (SC) and not in the inferior colliculus (IC, arrow) in control brains. In Smo-En1 cko embryos, Dmbx1 expression extends to the Cb, indicating a loss of the IC. Scale bars: 150 μm.