Anti-CD47 antibody inhibits chemokine-mediated migration of lymphoma but does not affect integrin-mediated adhesion. (A) Raji cells engineered with reduced CD47 expression levels (as in Figure 1) were assessed in a transwell migration assay with chemokines, either human SDF-1α (10nM) or CXCL13 (1 μg/mL) for 3 hours at 37°C. (B) Wild-type Raji cells were preincubated with the indicated antibodies at 10 μg/mL for 20 minutes and then assessed in a transwell migration assay for 3.5 hours at 37°C with the indicated chemokine. (A-B) The percentage of migrating cells relative to input was determined. Experiments were performed in duplicate and repeated. (C) Raji cell subclones were incubated on plates coated with human VCAM at 1 μg/mL unless indicated (VCAM negative = BSA-coated plates). (D) Raji cells were incubated in similar conditions to panel C with the indicated antibodies at 10 μg/mL. Adhesion of cells was determined by measuring absorbance at 490 nm, and relative absorbance to BSA-coated plates is shown ± SEM. No differences in VCAM-coated conditions were observed in either panel C or D. *P < .05 (t test). **P < .01 (t test). ***P < .005 (t test). ****P < .0001 (t test).