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Fig. 3. p53 negatively regulates neural stem cell proliferation, survival and
self-renewal. Neurospheres from the lateral ventricle wall of adult
p53-null mice grow faster and become larger compared with wild-type
neurospheres. (A,B) Secondary neurospheres 6 days after passage.
(C) Average volume of these neurospheres. (D) When neurospheres
are dissociated, a larger proportion of p53-/- compared
with wild-type cells are capable of forming new neurospheres, demonstrating an
increased self-renewal capacity. (E,F) Apoptotic cell death,
indicated by Annexin V (E) or caspase activity (F) detected by flow cytometry,
is decreased in the absence of p53. (G-I) A similar proportion
of neurospheres from p53 mutant and wild-type mice are tripotent and form
neurons (ßIII-tubulin+), astrocytes (Gfap+) and oligodendrocytes (O4+)
upon differentiation. (J,K) BrdU incorporation is increased in
p53-/- neurosphere cells compared with cells from
wild-type mice (K) and more cells from p53-/- mice have a
DNA content corresponding to S and M phases (J). There is no size difference
between wild-type and p53 mutant cells (L). All bars in graphs indicate
mean±s.d. *P<0.05,
**P<0.01, ***P<0.001. Scale bar:
100 µm.
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