Extended Data Figure 4. Anti-CD47 Ab promotes the phagocytosis of diseased SMCs and macrophages, without altering apoptosis.

(a). In vitro caspase activity assays reveal that anti-CD47 Ab does not alter rates of programmed cell death in any vascular cell type (b). Flow cytometry assays confirm that anti-CD47 Ab has no effect on apoptosis at baseline, or in vascular SMCs exposed to 24 or 72 hours of oxLDL. (c). Staining controls for the in vitro phagocytosis assays. (d). Representative FACS plots for the in vitro efferocytosis conditions displayed in Figure 2E. The right upper quadrant (highlighted in red) includes double positive cells which are taken to represent a macrophage that has ingested a target cell. (e). In vitro efferocytosis assays using lipid-loaded macrophages as the target cell confirm that anti-CD47 Ab also stimulates the clearance of this vascular cell type, similar to the findings observed with SMCs. (f). Additional in vitro efferocytosis assays confirm that anti-CD47 Ab stimulates phagocytosis of vascular cells in a specific manner. Error bars represent the SEM.