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Fig. 1. Fgf4-/- ES cells are deficient in neural
induction. (A-E) Immunostaining (left; phase-contrast, right) for
Oct4 and nestin (A,D) and Oct4 and TuJ1 (B,C,E) in wild-type (A,B),
Fgf4+/- (C) and Fgf4-/- mouse ES cells
on day 6 (A,D) and day 10 (B,C,E) of the monolayer neural differentiation
protocol. (F) RT-PCR for neural markers Sox1 and nestin in
Fgf4+/- and Fgf4-/- ES cells in
self-renewing conditions on day 2 of the neural induction protocol.
ß-actin was used as a loading control. (G) Quantitative
RT-PCR for nestin on day 5 of the neural differentiation assay. TBP,
TATA-binding protein. (H,I) Immunostaining for Oct4 and nestin
(H) and Oct4 and TuJ1 (I) of Fgf4-/- ES cells cultured in
FGF4 (5 ng/ml) for the first 24 hours of a 6 day (H) and 10 day (I) neural
monolayer assay. Scale bar: 100 µm. (J) Cell counts of
Oct4-positive, nestin-positive, and double-negative cells for wild-type,
Fgf4-/- ES cells, and Fgf4-/- ES cells
treated with FGF4 after 6 days culture in N2B27 (n=3 for each
sample).
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