Extended Data Fig. 6: Structure-function analyses of CD47 and SLAMF7.
From: CD47 masks pro-phagocytic ligands in cis on tumor cells to suppress antitumor immunity
a, Expression of mCD47 and mSLAMF7 and binding of mSIRPα-Fc (red lines) in L1210 derivatives assessed by flow cytometry. Filled curves, Ctrl IgG or Ctrl Fc fusion protein. b, Phagocytosis assays of L1210 derivatives by WT or Sirpa-/- BMDMs as in Fig. 2a. c, Expression of mSLAMF7 (red lines) in Slamf7-/- conA-activated CD4+ T cells, transduced with variants of mSLAMF7 constructs, assessed by flow cytometry. Filled curves, Ctrl IgG. d, Phagocytosis assays of conA-activated CD4+ T cells by WT or Sirpa-/- BMDMs as in Fig. 2a. e,f, Schematic representation (e) and time-course of normalized dye 505 fluorescence intensity (f) of LUV-based FRET assay of mSLAMF7 (donor) or hPD-L1 (donor) with mCD47 (acceptor) or hPD-1 (acceptor) monitored by a real time plate reader. g, Time-course of normalized dye 505 fluorescence intensity of LUV-based FRET assay of mSLAMF7 (donor) with mCD47 (acceptor), pre-treated with Ctrl IgG MOPC21, mSLAMF7 mAb 4G2 and mCD47 mAb Miap301, monitored by a real time plate reader. h, Binding of soluble hCD47-Fc fusion protein to WT or CD47-/- Raji cells assessed by flow cytometry. All data are means ± s.e.m. ns, not significant; **p < 0.01 and ***p < 0.001. Flow cytometry profiles are representative of 3 (a,c) or 2 (h) independent experiments. Results are representative of 3 (f) or 2 (g) independent experiments. Results are pooled from 4 (b) or 4 (d; for all variants, except 3 for SLAMF7R75A) independent experiments. Each symbol represents one mouse.
