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Fig. 7. Perturbation of endothelial-cell-monolayer integrity in co-culture spheroids of Ang-1 or Ang-2 overexpressing HUVECs and HUASMCs. Control HUVECs (B) or HUVECs retrovirally transduced with Ang-1 (C, endo. Ang-1) or Ang-2 (D, endo. Ang-2) were mixed with HUASMCs and allowed to form co-culture spheroids for 48 hours. Established co-culture spheroids were then kept in spinner culture for 4 hours and endothelial-cell surface coverage was quantified by automated image analysis following whole-mount staining for CD31 using Alexa Fluor 546 as fluorescent dye. Control HUVECs (B) and Ang-1 expressing HUVECs (C) form stable, differentiated co-culture spheroids with HUASMCs. By contrast, Ang-2 overexpressing HUVECs form co-culture spheroids with HUASMCs, with endothelial cells detaching upon mechanical challenge in spinner culture (A,D). Co-culture spheroids of Ang-2-overexpressing HUVECs and HUASMCs can be stabilized by exogenous Ang-1 (F) or VEGF (G) but not by sTie-2 (A,E). **, P<0.01 compared with control.





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