Fig. 1.

Characterization of c4D10 in vitro. (A, B) Antibodies were used to stain CHOK1/cynoCD47 cells (A, left) or CHOK1/hCD47 cells (A, right) or human RBCs (B) prior to detection with Alexa Fluor 488‐conjugated anti‐human secondary antibody by flow cytometry. The experiment was performed three times with similar results being obtained. (C, D) Macrophages were cocultured with CFSE‐labelled Jurkat cells in the presence of hIgG4, c4D10, CC‐90002 or Hu5F9‐G4. Representative cytofluorometric plots reflecting maximum phagocyticity are shown (C), the phagocytosis index was determined by the percentage of Alexa 488⁺ cells within the APC⁺ macrophage cell gate (D, left) and IC₅₀ values of indicated antibodies were calculated using prism, version 6 (D, right). Data shown represent n = 3 donors. (E) Competitive inhibition of SIRPα binding to CD47. Serial dilutions of c4D10, CC‐90002 and Hu5F9‐G4 disrupt the interaction of CD47 and SIRPα. Results are representative of three independent experiments. All error bars indicate the SEM.