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. 2000 Jan 10;148(1):203–216. doi: 10.1083/jcb.148.1.203

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© 2000 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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Inhibition of monocyte transmigration prevents monocyte-dependent changes in VE-cadherin staining. PBMC (10⁶/ml) were incubated in the presence of either binding control (W6/32 mAb, 10 μg/ml) or the combination of α4-integrin (mAb HP2.1) and β2-integrin (TS1/18 mAb, each at 10 μg/ml) for 15 min at room temperature. In additional experiments, PBMC and the HUVEC monolayer were incubated in the presence of either Ab 177 or preimmune IgG (20 μg/ml) for 30 min before use in flow assays. a, The total interacting cells (rolling, adherent, and transmigrated) were comparable in each case, 602 ± 157 cells/mm² in the presence of HP2.1 and TS1/18; 573 ± 53 cells/mm² in the presence of Ab177; and 575.8 ± 170 cells/mm² in the presence of preimmune IgG. Data are the mean ± SD, n = 4 (four coverslips analyzed per experiment). b, The number of monocytes associated with changes in VE-cadherin were quantified as described in Materials and Methods. Three fields were taken from each coverslip (two coverslips analyzed per experiment). Data are expressed as the mean ± SD, n = 4. The range of total leukocytes per field was 13–65 cells, with most fields falling between 25 and 47 cells.

Inhibition of monocyte transmigration prevents monocyte-dependent changes in VE-cadherin staining. PBMC (10⁶/ml) were incubated in the presence of either binding control (W6/32 mAb, 10 μg/ml) or the combination of α4-integrin (mAb HP2.1) and β2-integrin (TS1/18 mAb, each at 10 μg/ml) for 15 min at room temperature. In additional experiments, PBMC and the HUVEC monolayer were incubated in the presence of either Ab 177 or preimmune IgG (20 μg/ml) for 30 min before use in flow assays. a, The total interacting cells (rolling, adherent, and transmigrated) were comparable in each case, 602 ± 157 cells/mm² in the presence of HP2.1 and TS1/18; 573 ± 53 cells/mm² in the presence of Ab177; and 575.8 ± 170 cells/mm² in the presence of preimmune IgG. Data are the mean ± SD, n = 4 (four coverslips analyzed per experiment). b, The number of monocytes associated with changes in VE-cadherin were quantified as described in Materials and Methods. Three fields were taken from each coverslip (two coverslips analyzed per experiment). Data are expressed as the mean ± SD, n = 4. The range of total leukocytes per field was 13–65 cells, with most fields falling between 25 and 47 cells.