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Fig. 3. Requirement of FoxM1 for neuronal differentiation but not
specification. (A) Xenopus embryos pre-injected with
control MO or FoxM1-MO (36 ng) together with or without FoxM1 mRNA
(200 pg) at the one-cell stage were cultured until st. 33 and photographed
(left panels). For RT-PCR analysis, embryos were collected at st. 28 (right
panel). (B) Embryos injected with lacZ mRNA (100 pg) and
either control MO or FoxM1-MO (18 ng) at one blastomere at the two-cell stage
were cultured until st. 14 for WISH analysis of Xngnr1, N-tubulin and
MyoD, or until st. 16 for analysis of Sox2 and
N-CAM. In each panel, the injected side (β-Gal, red) of the
embryo is on the right (dorsal view, anterior up). (C) Animal caps from
the late blastula embryos pre-injected with control MO or FoxM1-MO (36 ng) at
the one-cell stage were cultured with or without Activin (200 pM) until
sibling control embryos reached st. 20 and then analyzed by RT-PCR. (D)
Animal caps pre-treated with Noggin mRNA (100 pg) and either control MO or
FoxM1-MO (36 ng) were cultured and analyzed as in C. (E) Animal caps
pre-treated or not with Noggin mRNA (100 pg) were incubated for the indicated
times and analyzed by RT-PCR. Whereas FoxM1 expression in control
animal caps decreased after 2 hours of incubation, that in Noggin-treated
animal caps remained constant until 12 hours of incubation, indicating that
the induction of FoxM1 expression by Noggin began after 2 hours of
incubation. Note that Sox2 began to be expressed coincidently with
FoxM1, whereas expression of N-CAM and N-tubulin began after
FoxM1, in Noggin-treated animal caps.
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