Immobilized P- or E-selectin triggers activated mouse T-cell death. ConA-stimulated activated mouse T cells were added to a 96-well plate precoated with (A) E-selectin or (B) P-selectin in titrated concentrations. After shear (80 rpm) for 30 minutes, the cells were incubated for a total of 5 hours. The cell death was assayed by cellular ELISA with biotinated Annexin, followed by streptavidin-β-gal. Color development by substrate ONPG was read at OD 420 nm. OD difference was shown as compared with BSA-coated wells. (C) Cell viability was assayed by fluorescence of Calcein am and analyzed by fluorescence microplate reader systems (excitation at 494 nm; emission at 530 nm). After incubation for 16 hours, the fluorescence of activated T cells in BSA-coated wells displayed at 756.4 and relative reduction in fluorescence of viable cells in wells coated with P- or E-selectins (5 μg/mL) compared with those in BSA-coated wells was shown. These data represent at least 3 independent experiments and were analyzed by general linear model with Bonferroni correction for multiple comparisons.