DEV02441 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 - Fig. S1. Fates of c2 and c3 populations. (A) At E13.5, early calbindin+ Purkinje cells are co-labeled with Lhx1/5+ in the c2 domain. A′ is a higher magnification of the boxed region. Arrowheads in A′ indicate double-labeled cells, arrow indicates a subpopulation of c2 cells that are calbindin negative. They may be newly born Purkinje cells that do not express calbindin yet or progenitors of Golgi cells or GABAergic DCN neurons. (B) Adult sagittal immunostained sections as marked. Arrowhead in B′ labels Purkinje cell; white arrowheads in B′′ show labeled stellate and basket cells; black arrowheads indicate Golgi cells. (B′′′) DCN cells. IGL, inner granule cell layer; PC, Purkinje cell layer; ML, molecular layer. (C) Lmx1a in situ indicates cells in the location of DCN (arrows). cpe, choroid plexus. (D) Sagittal section of E17.5 cerebellar anlage at the level of the vertical line in C. (D′) Double Lmx1a/calretinin immunohistochemistry of boxed region. Arrowheads indicate Lmx1a+/calretinin+ cells.

• Supplemental Figure 2 -

  Fig. S2. Lmx1a overexpression changes cell fates in the E16.5 cerebellum. (A,B) Hematoxylin and Eosin stained sagittal sections of the cerebellar primordium and (D-G) adjacent immunostained sections with markers and genotype indicated. The granule cells from c1 progenitors are increased in transgenic versus wild-type cerebella (C), while c2 cell fates are lost.

• Supplemental Figure 3 - Fig. S3. Cellular populations and morphology of dreher cerebellar anlage at E12.5. (A-D) Hematoxylin and Eosin stained paramedial (A,B) and lateral (C,D) sagittal sections at E12.5 showing altered morphology and lack of choroid plexus (arrowhead in A, C; asterisk in B, D) in dreher embryos. 3v and 4v, 3rd and 4th ventricles. (E-J) Immunohistochemistry and cell counts however indicate that all cell populations are present in dreher.

• Supplemental Figure 4 -

  Fig. S4. GDF7-DTA ablation is specific to the E9.5 roof plate. TUNEL-labeled transverse sections through r1 at E9.5 (A,B) and E10.5 (C,D) in wild-type (A,C) and roof plate-ablated embryos (B,D). Apoptotic cells (green dots) are specifically found in the most dorsal neuroepithelium (arrowhead) of roof plate-ablated embryos at E9.5. Insets show high magnification of boxed regions.