Forced α1-3-fucosylation of WEHI-3 cells with FTVI and GDP-fucose does not augment cell rolling on P-selectin or E-selectin in shear flow.
(A) WEHI-3 cells were perfused over immobilized murine P-selectin IgM chimera (90 sites/μm2) or E-selectin IgM chimera (140 sites/μm2) or control CD45 IgM chimera in the presence or absence of EDTA. After 4 minutes, the accumulated number of rolling cells was quantified by counting all cells in each of 4 randomly selected × 40 fields. The data represent the means ± SDs of 4 experiments. (B) Control or FTVI-treated WEHI-3 cells were perfused over P-selectin IgM (90 sites/μm2) or E-selectin IgM (140 sites/μm2) at 0.5 or 1 dyn/cm2. After 4 minutes, the accumulated number of rolling cells was quantified. The data represent the means ± SDs of 4 experiments. (C-D) Control or FTVI-treated cells were allowed to accumulate at 0.5 dyn/cm2, and cell-free buffer was then introduced. Wall shear stress was increased every 30 seconds, and the percentage of remaining adherent cells was determined. The data represent the means ± SDs from 3 experiments. (E-F) Control or FTVI-treated cells were perfused over P- or E-selectin IgM at 0.5 or 4 dyn/cm2. The data represent the means ± SDs for rolling velocities measured for 50 cells from 3 experiments.
