Supplemental Figure 1
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Fig. S1.
(A) Single cells from E14.5 cerebellar anlagen are able to proliferate and form
neurospheres in vitro. Neonatal (P0) and adult cerebellar cells maintain this
ability, although at reduced frequency. For comparison, cells from E14.5 medial
and lateral ganglionic eminences (GE) were also analyzed. (B) Cells from
GFP-positive and GFP-negative animals were mixed in equal proportion and plated
at what others have suggested are clonal densities. The developing neurospheres
were either GFP positive or GFP negative, and we never observed heterogenious
neurospheres containing both GFP positive and negative spheres, supporting the
clonality of our assay. (C) Differentiated neurospheres were doublestained with
GFAP and calbindin. GFAP-expressing cells never expressed calbindin making it
very unlikely that the calbindin expression in the neurospheres cultures is due
to reactive astrocytes. Doublestaining of secondary neurosphere confirm that
cerebellar derived-neurospheres maintain their cell fate potential and give
rise to neurons that resemble: granule cells, based on their expression of
glutamate (D) and TAG1 (E); Purkinje cells, based on their expression of
calbindin (D,E); and interneurons, based on their expression of GABA and
parvalbumin (F). Clonally derived adult cerebellar-derived neurospheres were
plated on feeder layers and gave rise to cells that resembled GABAerigic
interneurons (G), calbindin-expressing Purkinje cells (H) and glutamatergic
granule cells (I).
