|
|
|
|
|
|
|
||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | ||||
|
Fig. 3. Wnt and Notch signaling affect blast colony formation from the
hemangioblast. (A) Role of Wnt signaling in blast colony
development. Flk1+ cells (4x104) isolated from the
day 2.75 EBs generated from either sβ-cat, Numb or Notch1-IC ES cells
were cultured in M10 serum-free hemangioblast media in the presence or absence
of doxycyclin (Dox; 2 µg/ml) and DKK1(DKK; 300 ng/ml) as indicated. Dox was
added at 0 hours of culture, whereas DKK1 was added at 6 hours of culture.
Colonies were washed with media at 24 hours to deplete DKK1 and Dox, and
replated back into fresh M10. Blast and core colonies were scored after 4 days
of culture. (B) Effect of DKK1 treatment on blast and core colony
development. (C) Hematopoietic potential of DKK1-treated colonies.
Sixteen-hundred colonies of each group were picked at day 3 of culture,
pooled, dissociated and replated into hematopoietic methylcellulose cultures.
Ep, primitive erythroid colonies; Def, combined macrophage, bipotential
macrophage/erythroid, definitive erythroid and multipotential
myeloid/erythroid colonies. (D) Role of Numb in blast colony
development. gSI, 
-secretase inhibitor (2.5 µM). (E) Role of
Notch signaling in blast colony development. (A-E) Error bars represent
standard deviations of the mean of number of colonies from n
independent experiments (A, n=3; B, n=3; C, n=3; D,
n=4; E, n=4; **P<0.01;
***P<0.001)
|
