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Fig. 6. MMP-10 is induced during EC morphogenesis, is activated by serine proteases and treatment of ECs with siRNA targeting MMP-10 delays capillary tube regression and collagen gel contraction induced by multiple serine proteases. ECs were suspended in collagen matrices as described in the absence (A) or presence (B) of varying doses of plasma kallikrein. Gels were monitored for contraction (indicated by +) and conditioned media were collected for western blot analysis of MMP-10 at the indicated time points (A) or at 72 hours (B). Note that latent MMP-10 (arrows) is visible in each condition; however, active MMP-10 (arrowheads) is visible only in the presence of kallikrein. (C) ECs were transfected as described with MMP-1, MMP-10, MMP-2, MMP-9 or luciferase control siRNA and suspended in collagen matrices. At time zero, culture media was added with or without prekallikrein (2 µg/ml) and Factor XII (1 µg/ml) and percentage gel contraction was calculated over time. Conditioned media were collected and analyzed for MMP-10 or MMP-1 expression and activation, as well as for MMP-2 and MMP-9 (data not shown). Arrows and arrowheads indicate the latent or activated forms of MMPs, respectively.
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