Extended Data Fig. 4: Glutaminyl cyclase inhibition leads to reduced binding of recombinant hSIRPα.

a, Cell surface binding of αhCD47-2D3, αhCD47-CC2C6 and hSIRPα-Fc to control (DMSO)-treated (-) or SEN177-treated (+) lung cancer (A549), colorectal (DLD1), HAP1, rectal carcinoma (RKO) and breast cancer (SKBR3) cells, as determined by flow cytometry. Data represent n = 3 biological replicates and mean ± s.d. of triplicates. ***P≤0.000695 by unpaired two-sided t-test. b, Cell surface binding of αhCD47-2D3, αhCD47-CC2C6 and hSIRPα-Fc to control (DMSO)-treated (-), SEN177-treated, and PQ912-treated melanoma (A375) cells, as determined by flow cytometry. Data represent n = 3 biological replicates and mean ± s.d. of triplicates. ***P = 0.0001 by one-way ANOVA with multiple comparison correction. c, Flow cytometry plot of surface binding of αhCD47-B6H12 and αhCD47-CC2C6 to control-treated and PQ912-treated melanoma (A375) cells. Data are representative of two independent experiments with similar results (n = 3 biological replicates per experiment). d, Cell surface binding of αhCD47-2D3, αhCD47-CC2C6 and hSIRPα-Fc to control (DMSO)-treated (-) and SEN177-treated (+) wild-type and QPCTL-knockout epidermoid carcinoma (A431) and lung cancer (A549) cells, as determined by flow cytometry. Data represent n = 3 biological replicates and mean ± s.d. of triplicates. e, Cell surface binding of secondary antibody alone, hSIRPα-Fc (followed by secondary antibody) or hSIRPα-Fc in the presence of the CD47 blocking antibody αhCD47-B6H12 (followed by secondary antibody) to control (DMSO)-treated (-) or SEN177-treated lung cancer (A549) cells, as determined by flow cytometry. Data represent n = 3 biological replicates and mean ± s.d. of triplicates. Values indicate MFI relative to WT cells stained with the same reagent (a,b,d) or MFI (e). Data are representative of one (e) or at least two independent experiments (a–d).