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Fig. 3. Survival of MLNSC in insulin correlates with insulin-induced Akt
activation. Cell recovery and BrdU assays were performed on cultures of
CD31- CD35- EGFP+ neurospheres grown in
MB-media, MB-media minus insulin or MB-media plus 100 µM quercetin. (A)
Sphere recovery was determined by counting the number of spheres daily and
normalizing to the number of spheres grown in complete MB-media. Presented is
the mean±s.d. for three experiments from distinct cellular isolations.
(B) Proliferation is unaltered in BM-media. BrdU incorporation assays were
conducted on neurospheres grown in MB-media, MB-media minus insulin and
MB-media plus 100 µm quercetin. Little change in BrdU incorporation was
observed whether BrdU was delivered for 4 (B) or 8 (C) hours. Presented is the
mean±s.d. of three experiments normalized to neurospheres grown in
MB-media containing insulin. (D) Insulin-induced survival correlates with the
activation of Akt. Western analysis of neurospheres cultured in MB-media minus
insulin (lane A), in MB-media plus 100 µM quercetin (lane B) and in
MB-media (lanes C,D). (E) Caspase 3 is activated in neurospheres the absence
of insulin signaling. Western analysis of procaspase 3 and activated caspase 3
in MB-media (lanes A,B) or MB-media plus 100 µM quercetin (lane C).
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