American Society of Hematology

Figure 3.

$Figure 3. CD47-mediated adhesion of human RBCs to human SIRPα1ex-coated surfaces. (A) Human RBCs were allowed to adhere at modest density to SIRPα1excoated wells in a 96-well plate for 10 minutes and centrifuged inverted for 10 minutes at 100g (cells remained wet). (B) Microscopy showed significant, intact adhesion of human RBCs to SIRPα1ex-coated wells but minimal adhesion to GST. (C) The fraction of well area covered by adherent RBCs after centrifugation was very small for bovine serum albumin (BSA)- and glutathione S-transferase (GST)-compared with SIRPα1ex-coated surfaces. (D) Prelabeling of human RBCs with blocking antibodies (B6H12 and BRIC126) prevented cell adhesion, whereas 2D3 did not inhibit or significantly enhance adhesion. The G-forces used here exert a sustained peeling force of approximately 100 pN on a human red cell (volume = 95 fL; density = 1.09 g/mL). Error bars indicate plus or minus 1 SD from multiple experiments.$

CD47-mediated adhesion of human RBCs to human SIRPα1ex-coated surfaces. (A) Human RBCs were allowed to adhere at modest density to SIRPα1excoated wells in a 96-well plate for 10 minutes and centrifuged inverted for 10 minutes at 100g (cells remained wet). (B) Microscopy showed significant, intact adhesion of human RBCs to SIRPα1ex-coated wells but minimal adhesion to GST. (C) The fraction of well area covered by adherent RBCs after centrifugation was very small for bovine serum albumin (BSA)- and glutathione S-transferase (GST)-compared with SIRPα1ex-coated surfaces. (D) Prelabeling of human RBCs with blocking antibodies (B6H12 and BRIC126) prevented cell adhesion, whereas 2D3 did not inhibit or significantly enhance adhesion. The G-forces used here exert a sustained peeling force of approximately 100 pN on a human red cell (volume = 95 fL; density = 1.09 g/mL). Error bars indicate plus or minus 1 SD from multiple experiments.

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