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Soluble VCAM-1 binding to α4 integrins is cell-type specific and activation dependent and is disrupted during apoptosis in T cells by David M. Rose, Pina M. Cardarelli, Ronald R. Cobb, and Mark H. Ginsberg Blood Volume 95(2):602-609 January 15, 2000 ©2000 by American Society of Hematology sVCAM-1 binding to isolated peripheral blood lymphocytes.Human peripheral blood lymphoid cells were isolated as described in “Materials and Methods.” Cells were resuspended in Tyrode's buffer containing 1 mmol/L CaCl2, 1 mmol/L MgCl2, and 0.1% BSA and incub... David M. Rose et al. Blood 2000;95:602-609 ©2000 by American Society of Hematology PMA stimulates sVCAM-1 binding to CD45RO-positive peripheral T-lymphocytes.Human peripheral blood T-lymphocytes were isolated as described in “Materials and Methods”. David M. Rose et al. Blood 2000;95:602-609 ©2000 by American Society of Hematology CD16-positive human peripheral blood natural killer cells constitutively bind sVCAM-1.Human peripheral blood lymphoid cells were isolated as described in “Materials and Methods”. David M. Rose et al. Blood 2000;95:602-609 ©2000 by American Society of Hematology David M. Rose et al. Blood 2000;95:602-609 ©2000 by American Society of Hematology David M. Rose et al. Blood 2000;95:602-609 ©2000 by American Society of Hematology David M. Rose et al. Blood 2000;95:602-609 ©2000 by American Society of Hematology Cell-type–specific binding of sVCAM-1 to 4β1 integrin.Jurkat, CHO-α4β1, RBL, THP-1, U937, or HUT-78 was resuspended in either DMEM containing 0.1% BSA (solid line), DMEM with the addition of 5 mmol/L EDTA (shaded line), or Tyrode's buffer containing 1 mmol... David M. Rose et al. Blood 2000;95:602-609 ©2000 by American Society of Hematology Cellular-energy depletion suppresses sVCAM-Ig binding.Jurkat (A) or CHO-α4β1 (B) cells were placed in glucose-free Tyrode's buffer containing 2-deoxyglucose (2 mg/mL) and sodium azide (0.1% wt/vol) and were incubated at 37°C for 0, 30, 60, or 120 minutes. David M. Rose et al. Blood 2000;95:602-609 ©2000 by American Society of Hematology Fas-induced apoptosis suppresses sVCAM-Ig binding to Jurkat cells.Jurkat cells were untreated (A) or were treated with anti-Fas antibody CH11 (300 ng/mL) for 4 hours (B-D). 7 Ig-domain VCAM-Ig binding was assayed as described in “Materials and Methods” (A-C). David M. Rose et al. Blood 2000;95:602-609 ©2000 by American Society of Hematology Activated H-Ras and Raf-1 suppress 4β1 integrin-mediated VCAM-1 ligand binding.CHO-α4β1 cells were transiently transfected with cDNA encoding Tac-α5 or Tac-β1 alone or with a combination of Tac-α5 plus vector control DNA, H-Ras G12V, or RafBxB. David M. Rose et al. Blood 2000;95:602-609 ©2000 by American Society of Hematology