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Fig. 7. Ectopic expression of ZBP-89 impairs angiogenesis in mouse EB
cultures. (A) Primary day 11 EBs were dissociated and cultured in
collagen matrix with growth factors for endothelial cell differentiation for
four days. Histograms (mean±s.d., n=3) show the number of
primary day 11 EBs with 0, <5 or >5 branching vessels/EB derived from
the D5 and E1 clones, compared with the control.
*P<0.01; **P<0.001; NS, not
significant. (B) Representative images of control-, E1- or D5-derived
sprouting EB colonies. The reduction in the branching vessels (arrows in
controls) in EBs derived from overexpressing ZBP-89 clones E1 and D5 is also
clearly reflected by the compactness of the respective colonies; fewer
branches are produced compared with the controls. (C,D) Flow
cytometric analysis of a single-cell suspension of EBs overexpressing ZBP-89
and stained with PE-labeled anti-CD31 (day 6 culture) or anti-VE-Cadherin (day
7 and day 10 cultures) antibodies. ZBP-89 overexpression significantly reduced
the number of CD31+ and VE-Cadherin+ angioblasts
(similar results were obtained in two other independent experiments).
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