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Fig. 7. Soluble factors can restore mesoderm generation in the presence of
Dkk1. (A) ES cells were differentiated in SCM (no treatment), SCM
with Dkk1 alone, or in SCM + Dkk1 and one of the following factors:
TGFß1, Bmp2, Bmp4, Nodal, Fgf8, cripto or FST. On day 4 of
differentiation, cells were examined for Flk1 expression by FACS. Numbers
indicate the percentage of Flk1+ cells in the indicated region.
(B) ES cells were treated as described in A, and on day 4, embryoid
bodies were washed, cultured for an additional 2 days in SCM and then analyzed
for hematopoeitic potential as described in
Fig. 5A. (C) Cells were
treated as described in A, embryoid bodies washed and transferred to
gelatinized dishes in serum-free medium for 4 days. Cultures were analyzed at
day 8 for cardiac gene expression by RT-PCR. (D) ES cells were
differentiated in SCM alone (SCM) or SCM + Dkk1 in the absence (-) or presence
of Bmp4 or cripto, as indicated. Bmp4 and cripto were added on day 1, and
cells were analyzed on days 3 and 4 as indicated.
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