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Fig. 4. Combined effects of FGF-2 and VEGF-A. (A) EBs were cultured in the presence of individual factors VEGF-A (80 ng/ml) or FGF-2 (80 ng/ml) or a combination of the two (20 ng/ml FGF-2 and 30 ng/ml VEGF-A) as indicated from day 0 to day 8, followed by immunohistochemical anti-CD31 staining. Scale bars: 100 µm (upper) and 200 µm (lower). Boxed regions are shown at a higher magnification in the lower panels. (B) Western blotting of total lysates of 8-day EBs showed growth factor-regulated expression of VEGFR-2, CD31 and VE-cadherin. ß-catenin was used as loading control. B, basal; F20, FGF-2 at 20 ng/ml; V30, VEGF-A at 30 ng/ml; F20+V30, FGF-2 at 20 ng/ml and VEGF-A at 30 ng/ml). (C) Quantification of protein bands in western blot shown in Fig. 4B.





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