FIG E2.

Ligation-induced Siglec-8 endocytosis is not unique to the 2C4 mAb clone. Eosinophils were incubated with anti–Siglec-8 (either clone 2C4 [black columns] or 1H10 [white columns]) for 20 minutes at 4°C. Excess antibody was washed off, and the cells were incubated at 37°C for the indicated durations to permit endocytosis. Antibody remaining at the surface of cell was detected with a secondary antimouse IgG1 antibody. Data represent the means ± SDs of 4 independent experiments. ***P < .001; ****P < .0001 relative to eosinophils incubated with 2C4 mAb without any incubation at 37°C. ##P < .01; ###P < .001; ####P < .0001 relative to eosinophils incubated with 1H10 mAb without any incubation at 37°C.