FIGURE 2.

MMP-2 mediates 15(S)-HETE-induced HDMVEC migration and tube formation in vitro and Matrigel plug angiogenesis in vivo. A and B, quiescent HDMVECs were treated with and without 10 mm GM6001 for 30 min at 37 °C, trypsinized, rinsed with trypsin-neutralizing solution, and subjected to 0.1 μm 15(S)-HETE-induced migration (A) or tube formation (B). C, WT mice were injected subcutaneously with 0.5 ml of Matrigel premixed with vehicle or 10 μm 15(S)-HETE with and without 10 mm GM6001. One week later, the animals were sacrificed, and the Matrigel plugs were harvested from underneath the skin, and either cryosections were made and examined by double immunofluorescence staining for vWF and CD31 using their specific antibodies or analyzed for hemoglobin content using Drabkin's reagent. D, HDMVECs were transfected with scrambled or MMP-2 siRNA, and 48 h later cell extracts were prepared and analyzed by Western blotting for MMP-2 levels using its specific antibodies. E and F, all the conditions were the same as in D except that, after transfection, HDMVECs were quiesced and subjected to 15(S)-HETE (0.1 μm)-induced migration (E) or tube formation (F). The bar graphs in A–F represent the mean ± S.D. values of three independent experiments or six plugs from six animals. *, p < 0.01 versus control; **, p < 0.01 versus 15(S)-HETE. TR, transfection reagent.