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. 2008 Feb;172(2):395–405. doi: 10.2353/ajpath.2008.070870

Figure 6.

Figure 6

WT and galectin-3−/− mouse neutrophils undergo apoptosis at similar rates in culture, and galectin-3 protects human neutrophils from apoptosis. A: Mouse bone marrow neutrophils were cultured for the indicated time points and incubated with PE-conjugated anti-GR-1 antibody followed by annexin-V-FITC. ToPro-3 was added to each sample before FACS analysis. Apoptotic and necrotic neutrophils were identified as those demonstrating GR-1 and annexin-V/ToPro-3 positivity (n = 3). B: Human peripheral blood neutrophils were cultured for 18 hours in Iscove’s modified Dulbecco’s medium containing 10% FBS and 30 μg/ml of human recombinant galectin-3 or PBS. Cytospin preparations were prepared, and neutrophil apoptosis was determined by morphology. Arrows show apoptotic human neutrophils. Percent apoptosis was calculated from total cell number of five fields of each slide (n = 3) (**P < 0.01 compared to untreated). C: Human peripheral blood neutrophils were cultured for 18 hours in Iscove’s modified Dulbecco’s medium containing 10% FBS and 20 μg/ml of human recombinant galectin-3. Neutrophils were incubated with annexin-V-FITC, and ToPro-3 was added to each sample before FACS analysis. Annexin-V-positive and ToPro-3-negative neutrophils were classed as apoptotic (n = 3) (*P < 0.05 compared to untreated).