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. Author manuscript; available in PMC: 2019 Apr 2.
Published in final edited form as: J Allergy Clin Immunol. 2017 Jul 20;141(5):1774–1785.e7. doi: 10.1016/j.jaci.2017.06.028

FIG 6.

FIG 6.

A toxin-conjugated mAb targeted to Siglec-8 causes cell death of eosinophils and a malignant mast cell line in vitro. A and B, Eosinophils were primed with 15 ng/mL IL-5 or left unprimed and concurrently treated with equimolar amounts of anti–Siglec-8 mAb (anti-Sig8), isotype control mAb, saporin (SAP)-conjugated 2C4, or saporin-conjugated isotype control mAb or were left untreated for 18 to 24 hours. C and D, HMC-1.2 cells were preincubated with the indicated mAbs or conjugates, washed extensively, and incubated for 7 to 13 days. Cell death was measured using FITC-labeled annexin V and DAPI staining and flow cytometry. Fig 6, A and C, Representative flow plots for each treatment. Percentages of live cells are shown. Fig 6, B and D, Percentages of viable (annexin V–negative, DAPI-negative) cells for each treatment normalized to the untreated group for each priming condition. Ab, Antibody; FITC, fluorescein isothiocyanate. Error bars represent the means ± SDs (B and D) of 12 and 8 independent experiments, respectively. *P < .05, ***P < .001, and ****P < .0001.