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Fig. 7. Augmented endothelial generation by GATA2. (A) GATA2
induction scheme in differentiating EBs and formation of endothelial cells.
Uppermost numbers indicate EB day. Gray bar indicates duration of Dox
treatment and subsequent analyses of D6 EBs. (B) Generation of
Tie2+ endothelial cells by GATA2 expression in serum and SR
conditions. Numbers in boxes indicate percentage of Tie2+ cells.
(C) Generation of CD31+ and
CD31+VE-Cadherin+ endothelial cells in SR media. Numbers
in upper left and right quadrants indicate percentage of CD31+ and
CD31+VE-Cadherin+ cells, respectively. (C')
Quantification of CD31+ and CD31+VE-Cadherin+
cells generated with or without Dox. Values are mean±s.e.m. from four
independent experiments; *, P<0.001 versus untreated.
(D-E') Representative sprouting EB images. D and E show
100x magnification of brightfield images of EBs generated in the absence
(D) and presence (E) of Dox; D' and E' are 800x
magnifications of D and E, respectively, showing the tendril-like (D')
and endothelial (E') cells generated in the two conditions.
(F,F') Brightfield (F) and fluorescence (F') images
of 800x magnification of sprouting iGATA2 EBs demonstrating uptake of
DiL-Ac-LDL by endothelial cells. (G) Fractional quantification of types
of sprouting structures grown by 44 (-Dox) and 46 (+Dox) EBs grown in
sprouting media.
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