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Fig. 5. Treatment of ECs with MMP-1 siRNA decreases MMP-1 protein expression and delays capillary tube regression and collagen gel contraction. ECs were transfected as described with MMP-1, 
2 macroglobulin (A2Mac), or luciferase control siRNA duplexes at a final concentration of 200 nM, suspended in collagen matrices and placed in the quantitative 384 micro-well regression assay (as in Fig. 3). At time zero, culture media was added with or without serine proteases at the doses indicated and the percentage of gel contraction was calculated over time. (A) Culture media contained control media, plasma kallikrein, or prekallikrein and Factor XII at the indicated doses. Upon completion of gel contraction, conditioned media were collected and analyzed for MMP-1 expression and activation. (B) ECs transfected with the indicated siRNAs were established in collagen matrices for 46 hours in the absence of serine proteases and were fixed, stained and photographed. Arrows denote newly established EC vascular networks and open lumenal structures. Bar, 100 µm.
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