Confocal imaging of adherent GD and CTR RBCs to laminin α5 at different shear stresses. (A) Typical microscopic images showing the shape of adherent RBCs from 7 healthy CTRs and 7 GD patients at 1 dyn/cm². The flow is from the left to the right. Lu/BCAM cell-surface expression was visualized by immunofluorescence using the monoclonal antibody F241 and confocal microscopy. Percentage of adherent RBCs with racket shape after inflow at 0.2 dyn/cm² and increasing washout until 1dyn/cm² was higher in CTR RBCs compared with GD RBCs (P < .01, Mann-Whitney test). (B) Same experiments as in panel A but with final washout at 4 dyn/cm². GD RBCs frequently exhibited typical tether shape. Bars scales represent 1 μm. RBCs from GD patients exhibited more frequently a tether shape than CTR RBCs (***P < .0001). RBCs from GD patients exhibited longer tethers than CTR RBCs. Tether measurement was performed on 30 cells per condition (***P < .0004). (C) Same experiments as in panel B but GD RBCs were double-stained with anti-Lu/BCAM (top panel) or anti-spectrin (bottom panel) antibodies.