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Figure 4


Fig. 4. Inactivation of Tgfbr2 in endocardial cells prevents EMT in vitro. (A,B) In vitro collagen gel assays performed with AV explants isolated from control (A) or mutant (B) embryonic hearts at E9.25. (C) The number of mesenchymal cells formed in control (Tgfbr2loxp/loxp;R26R) explants (n=18) is dramatically higher than that in mutant explants (n=14). *P<0.005. (D-G) Control (D,F) and mutant (E,G) explant cultures stained with an antibody (green) against the panendothelium marker Pecam (D,E), or against the mesenchyme marker {alpha}-smooth-muscle actin (F,G). Nuclei were visualized by staining with propidiumiodide (red). In the control, the mesenchymal cells expressed {alpha}-smooth-muscle actin (F) and lost the expression of Pecam (D). By contrast, the endocardial cells in mutant explant cultures expand on the gel surface with the expression of Pecam (E) but not of {alpha}-smooth-muscle actin (G). cko, Tie2cre;Tgfbr2loxp/loxp;R26R embryos; ctrl, control embryos.





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