DEV02437 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Distribution of embryonic cerebellar markers in E15.5 Ebf2 null mutant versus wt cerebellum. All sections are frontal. (A,C,E,G) In the wt Ebf1-Ebf3 are expressed similarly to RORa. In the mutant, the medial expression domain of Ebf1 is partially lost in the middle third of the developing hemicerebellum (arrows in A,B) and a conspicuous number of PCs expressing Ebf3 and RORa are malpositioned (arrowheads in F,H). In addition, beta-galactosidase- and Ebf3-positive PCs are both associated with PCs abnormally retained in the CTZ, suggesting a delay in their migration (arrows in D,F). Reelin mRNA distribution is unchanged in wt versus null mutant cerebellum (arrows and arrowheads in I,J). (A-C,E-J) Non-radioactive in situ hybridization of 14 μm frozen sections; (D) Anti-beta-galactosidase immunostaining. A,C,E,G,I are adjacent wt sections; B,F,H,J are adjacent mutant sections. Mes: mesencephalon; m,i,l: medial, intermediate and lateral domains, respectively. Scale bar: in H, 100 μm for A-H. The results are representative of three experiments, each conducted in a different litter, comparing one mutant E15.5 embryo and one wild type littermate.

• Supplemental Figure 2 -

  Fig. S2. Expression of PC striped markers in the postnatal cerebellar hemispheres. Non-radioactive in situ hybridization of 20 μm hemicerebellar frontal sections at P6. (B) The distribution of PCs expressing Pcdh10 is altered in the mutant hemisphere (arrows) while the total number of PCs expressing Pcdh10 is unchanged. (D) The loss of a large number of PCs is revealed by the Sema3A riboprobe. The expression of both markers in the DCN appears normal. (A,C) wt controls in A and C, respectively. The expression of both markers in the DCN appears normal. f: fastigial nucleus; d: dentate nucleus. Scale bar: in D, 200 μm for A-D.