Specialized 96 well e plates

Rhesus macaque PBMC, T cell, NK cell and macrophage killing assays were performed on the xCELLigence MP platform (ACEA Biosciences). Specialized 96-well E-plates (ACEA Biosciences) were coated with collagen (Sigma-Aldrich), and 4 × 10⁴ target iPSCs were plated in 100 μl of media. After the cell index reached 0.7, the effector cells were added at an E:T ratio of 1:1. NK cells were stimulated with 1 μg ml⁻¹ human IL-2 (PeproTech). As killing control, cells were treated with 2% Triton X-100 in water. Data were standardized and analyzed with RTCA 2.1 software (ACEA Biosciences).

NK cell and macrophage killing assays were performed on the XCelligence MP platform (ACEA BioSciences, San Diego, CA.). Specialized 96-well E-plates (ACEA BioSciences) were coated and 4 × 10⁴ target T cells were plated in 100 μl T cell media. After the Cell Index reached 0.7, effector cells were added at an effector cell to target cell (E:T) ratio of 1:1. NK cells were stimulated with 1 μg /ml human IL-2 (Peprotech). In some wells, an anti-CD47 fusion protein (IgG1 and IgG4, Creative Biolabs) or SIRPa-polyHis protein (Arco Biosystems) were added at a concentration of 100 μg/ml. As killing control, cells were treated with 2% TritonX100 or media only was added (Supplementary Fig. 15). Data were standardized and analyzed with the RTCA software (ACEA).

NK cell killing assays were performed on the xCELLigence MP platform (ACEA Biosciences). Specialized 96-well E-plates (ACEA Biosciences) were coated with tumor-coating solution (Agilent), and 4 × 10⁴ target K562 were plated in 100 μl of media. After the cell index reached 0.7, the effector cells were added at an effector cell to target cell (E:T) ratio of 1:1. NK cells were stimulated with 1 μg ml⁻¹ human IL-2 (PeproTech). As killing control, cells were treated with 2% Triton X-100 in water. Data were standardized and analyzed with RTCA 2.1 software (ACEA Biosciences). Supernatants were collected for granzyme B and perforin ELISA after 90 h.