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Figure 5


Fig. 5. Forced expression of integrin β3 in integrin-β1-null EBs rescues endoderm morphogenesis. (A) EBs were cultured for 5 days using integrin-β1-null ES cells stably transfected with integrin-β3–GFP (β3-GFP) or GFP. The expression of integrin β3 was analyzed by western blotting. (B) Wild-type and integrin-β1-null EBs expressing integrin-β3–GFP or GFP were cultured for 5 days and endoderm formation was examined by live phase microscopy. The percentage of EBs with endoderm formation was plotted. n=344 for the wild-type, n=328 for the GFP and n=311 for the β3-GFP group. (C) The β3-GFP or GFP EBs were cultured for 5 days and immunostained for integrin β3 and the endoderm markers Dab2 and GATA4. BM was stained for laminin {alpha}1 or perlecan. Integrin-β3–GFP is recruited to the BM zone (arrowheads). The inset is a 2x magnification of the boxed region to show GATA4 nuclear staining. en, endoderm; epi, epiblast.





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