Figure 3.
Figure 3. Examination of microvascular ECs. (A-C) Flow cytometric analysis of fusion protein expression in microvascular cardiac ECs (A), platelets (B), or monocytes (C) from the 2 transgenic strains. Open profiles indicate antihirudin or anti-TFPI mAb; gray shaded profiles, isotype-matched control mAb; ordinate, cell number; abscissa, intensity of fluorescence. (A) EC phenotype confirmed by expression of CD31 and anti-CD105. Activation induced in vitro by incubation with 1 μM PMA for 30 minutes. (B) Platelet phenotype confirmed by expression of CD41. Activation induced in vitro by incubation with 1 U/mL thrombin for 30 minutes. (C) Monocyte phenotype confirmed by expression of CD14. Resting monocytes purified from saline-treated mice. Activated monocytes purified from LPS/l-NAME–treated mice. (D) Clotting of recalcified mouse plasma in the presence of ECs from B6, Hir-Tg, or TFPI-Tg mice. Comparing results using resting ECs (open bar) from B6 with those using resting ECs from either transgenic strain, P = NS. Comparing results using IL-1α/PMA-activated ECs (filled bars) from B6 with those using similar ECs from either of the transgenic strains, P < .0001. Error bars have been included but are too small to see.

Examination of microvascular ECs. (A-C) Flow cytometric analysis of fusion protein expression in microvascular cardiac ECs (A), platelets (B), or monocytes (C) from the 2 transgenic strains. Open profiles indicate antihirudin or anti-TFPI mAb; gray shaded profiles, isotype-matched control mAb; ordinate, cell number; abscissa, intensity of fluorescence. (A) EC phenotype confirmed by expression of CD31 and anti-CD105. Activation induced in vitro by incubation with 1 μM PMA for 30 minutes. (B) Platelet phenotype confirmed by expression of CD41. Activation induced in vitro by incubation with 1 U/mL thrombin for 30 minutes. (C) Monocyte phenotype confirmed by expression of CD14. Resting monocytes purified from saline-treated mice. Activated monocytes purified from LPS/l-NAME–treated mice. (D) Clotting of recalcified mouse plasma in the presence of ECs from B6, Hir-Tg, or TFPI-Tg mice. Comparing results using resting ECs (open bar) from B6 with those using resting ECs from either transgenic strain, P = NS. Comparing results using IL-1α/PMA-activated ECs (filled bars) from B6 with those using similar ECs from either of the transgenic strains, P < .0001. Error bars have been included but are too small to see.

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