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. 2018 Jul 9;9:1542. doi: 10.3389/fimmu.2018.01542

Figure 4.

Figure 4

Vesicular trafficking of IFN-γ in the cytoplasm of human eosinophils. IFN-γ-positive Eosinophil Sombrero Vesicles (EoSVs) [highlighted in pink in (Ai) and (Bi)] are seen in activated cells after stimulation with tumor necrosis factor alpha (TNF-α) (A) or CCL11 (B). Note that labeling is mostly associated with the vesicle membranes. EoSVs carrying IFN-γ are distributed in the peripheral cytoplasm (A) and in close association with secretory granules (Gr) undergoing release of their contents (A,B). (C) After stimulation, the total numbers of IFN-γ-positive EoSVs significantly increased. (D) Most EoSVs immunolabeled for IFN-γ were observed in the cell periphery. Arrowhead in (A) indicates extracellular release of IFN-γ. Data represent mean ± SEM. NS, not stimulated. The numbers of labeled and not labeled EoSVs (n = 1,357 vesicles) were counted in cell sections (n = 30). ##P = 0.0051 versus NS group; +++P = 0.0004 versus NS group. Cells were isolated from the peripheral blood, stimulated or not with CCL11 or TNF-α, and prepared for pre-embedding immunanogold EM.