Figure 3.

CD11b-deficient neutrophils exhibit impaired adhesion but increased activity upon Mincle activation. (a) Bone marrow (BM) neutrophils from WT and CD11b-deficient mice were treated with 50 μg ml⁻¹ TDM, and the surface expression of CD11b and CD18 was determined by flow cytometry 24 h after stimulation. (b) WT and CD11b^−/− BM neutrophils were prelabeled with calcein acetoxymethyl ester 6 h before treatment with 50 μg ml⁻¹ TDM treatment, with or without priming with 10 ng ml⁻¹ IFN-γ (I+T). Adhered cells were imaged with a fluorescent microscope (bottom panel, × 40). The percentage of adherent cells was determined by measuring the fluorescence with a microplate reader at 492 nm before and after washing (b, top panel). (c) IL-6 and TNF-α levels were assayed 24 h after TDM stimulation. Data are representative of three independent experiments. *P<0.05, **P<0.01, ***P<0.0001 (two-tailed unpaired Student’s t-test).