Figure 6.
Leukocyte-endothelial interactions. (A) HUVECs were treated with poly(I:C) (12.5 μg/mL) and thrombin (5 nM) for 3 hours, and the abundance of E-selectin and VCAM-1 mRNA relative to GAPDH was measured by quantitative RT-PCR. (B) Representative scatterplots depicting cell surface expression of VCAM-1 and E-selection determined by flow cytometry in HUVECs treated for 4 hours with poly(I:C) and/or thrombin. (C-D) Representative micrographs of static adhesion assay of rhodamine B-labeled THP-1 cells to HUVECs treated with poly(I:C) and/or thrombin, vorapaxar, GB83, or anti-E-selectin– and anti-VCAM1 blocking antibodies (10 μg/mL) and respective nonimmune IgG controls (10 μg/mL). Original magnification, ×20. Brightness adjusted to 75% with Powerpoint picture correction. Bars represent 200 µm. (D) The mean ± standard deviation of adherent THP-1 cells per visual field from 2 independent experiments (n ≥ 9 for control, thrombin, poly(I:C); n ≥ 6 for anti-VCAM-1/E-selectin). *P < .05; **P < .01; ***P < .001; ****P < .0001 by ANOVA, followed by multiple comparison test.

Leukocyte-endothelial interactions. (A) HUVECs were treated with poly(I:C) (12.5 μg/mL) and thrombin (5 nM) for 3 hours, and the abundance of E-selectin and VCAM-1 mRNA relative to GAPDH was measured by quantitative RT-PCR. (B) Representative scatterplots depicting cell surface expression of VCAM-1 and E-selection determined by flow cytometry in HUVECs treated for 4 hours with poly(I:C) and/or thrombin. (C-D) Representative micrographs of static adhesion assay of rhodamine B-labeled THP-1 cells to HUVECs treated with poly(I:C) and/or thrombin, vorapaxar, GB83, or anti-E-selectin– and anti-VCAM1 blocking antibodies (10 μg/mL) and respective nonimmune IgG controls (10 μg/mL). Original magnification, ×20. Brightness adjusted to 75% with Powerpoint picture correction. Bars represent 200 µm. (D) The mean ± standard deviation of adherent THP-1 cells per visual field from 2 independent experiments (n ≥ 9 for control, thrombin, poly(I:C); n ≥ 6 for anti-VCAM-1/E-selectin). *P < .05; **P < .01; ***P < .001; ****P < .0001 by ANOVA, followed by multiple comparison test.

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