Figure 6
Cell adhesion to recombinant E-selectin-IgG under static and fluid shear conditions.
(a) Human E-selectin-IgG binding to WT and KO cells measured using flow cytometry (Mean Fluorescence Intensity). P2H3 is an anti-E-selectin blocking mAb. * and † P < 0.05 with respect to all other treatments except bars indicated by these symbols are not different from each other. Error bars are too small to be visible in some cases. (b) WT and KO cells interacting with immobilized E-selectin-IgG in microfluidic flow cells at a wall shear stress of 1 dyn/cm2. Cells containing truncated N- and O-glycans (i.e. [ON]− cells) were not captured from flow. * and † P < 0.05 for rolling and adherent cells respectively, with respect to WT cells. Statistics are not presented for anti-E-selectin blocking studies since cell adhesion was negligible in all cases. (c) Different cell types were captured under no-flow/static conditions for 2 min. prior to ramp-increase in wall shear stress starting with 1 dyn/cm2 between 0–2 min. Detailed shear protocol is shown on right axis using dashed line. (d) Representative figure from experiment in panel c at 0 and 15 s. [NOG]− cells were released immediately upon initiation of flow. [ON]−and WT cells continued to roll for longer times.
