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Fig. 7. Shh regulates proliferation and neurosphere formation in cooperation with
EGF. (A) Image of neurospheres from adult SVZ cultures. (B) Quantification of
the number of primary neurospheres formed from cultures of SVZ cells
previously grown on astrocytic monolayers with or without exogenous Shh (5
nM). Shh was not added to the neurosphere cultures. (C) Synergism of Shh and
EGF on neurosphere proliferation. The assay was done with a constant dose of
EGF at 1 ng/ml, and varying doses of Shh at 5 or 0.5 nM (left) or with a
constant does of Shh at 5 nM and varying doses of EGF at 5 and 0.5 ng/ml
(right). Treatments were for 48 hours. (D,E) Quantification of proliferation
as measured by the percentage of BrdU+ cells (D) and the number of
clones obtained in cloning assays (E) in adult SVZ neurospheres treated with
cyclopamine (5 µM) or treated with an equal dose of ethanol used as carrier
for in vitro work. In all cases, error bars indicate s.e.m. of triplicate
cultures.
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