Characterization of neutrophil-DC hybrids purified from crude BM culture. (A) BM cells from C57BL/6 mice were cultured with GM-CSF, Flt3L, or no added growth factor for the indicated periods, and resulting populations were examined for the expression of CD11c, Ly6G, and B220. The data show the numbers of total cells, CD11c⁺/Ly6G⁻ DCs, Ly6G⁺/CD11c⁺ neutrophil-DC hybrids, and B220⁺/CD11c⁺ pDCs (means ± SD from triplicate cultures). (B-E) Ly6G⁺/CD11c⁻/MHC II⁻ neutrophils, Ly6G⁺/CD11c⁺/MHC II⁺ neutrophil-DC hybrids, and Ly6G⁻/CD11c⁺/MHC II⁺ traditional DCs were purified on d 6 from GM-CSF–supplemented BM culture. These populations were examined under light microscopy after HEMA-3 staining (B), under differential interference contrast microscopy (C), or under confocal microscopy after staining with phalloidin and antivinculin mAb (D) or with anti-MPO mAb (E). All the images are representative of at least 3 independent experiments (bar represents 20 μm). Time-lapse images recorded under differential interference contrast microscope are shown in supplemental Video 1.