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Fig. 4. Erk2-/- ES cells are severely deficient in neural
and mesodermal commitment. (A) Immunoblotting for phospho-Erk1/2
(pErk1/2) and total Erk1/2 from serum-starved Erk2+/- and
Erk2-/- mouse ES cells that were stimulated with foetal
bovine serum (FBS), LIF, or unstimulated (-). (B-D) Immunostaining
(left; phase-contrast, right) for nestin in Erk2+/- cells
(A), and two independent Erk2-/- ES cell clones (B,C)
fixed on day 6 of neural monolayer culture. Scale bar: 50 µm. (E)
RT-PCR for Oct4, Fgf5, Rex1 and ß-actin (as loading
control) on RNA isolated from two Erk2-/- ES cell lines
(B1, B3) that were maintained in N2B27 medium without LIF. Parallel cultures
were exposed to LIF for 2 days (+LIF) before isolation of RNA. (F)
RT-PCR analysis of wild-type and two Erk2-/- (B1, B3) ES
cell lines on day 4 of mesoderm monolayer differentiation. (G) FACS
analysis for Pdgfr
expression of wild-type and
Erk2-/- ES cells on day 5 of mesoderm monolayer
differentiation.
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