Fig. 5: Necroptotic EVs modulate cytokine and chemokine secretion.

A Schematic overview of EV phagocytosis assay. EVs from untreated (none), apoptotic (TB), or necroptotic (TBQ) CFSE-stained U937 cells were added to TG-macs overnight. B TG-macs were then harvested and analyzed by flow cytometry for their CFSE positivity, demonstrating uptake of the EVs. C Supernatants were collected, and secreted cytokine and chemokine levels were analyzed by ELISA. B Data are presented as the mean of triplicate samples ±SD. ^*p < 0.05, ^**p < 0.01, ^****p < 0.0001 (one-way ANOVA followed by Tukey’s multiple-comparison test). C Empty symbols are triplicate samples of mouse#1, colored symbols are triplicate samples of mouse#2. Data are presented as the mean of duplicate mice ±SD, ^*p < 0.05, ^**p < 0.01, ^****p < 0.0001 (two-way ANOVA followed by Tukey’s multiple-comparison test), and are representative of two independent experiment, n = 5. TG-macs, thioglycolate-derived peritoneal macrophages; EVs, extracellular vesicles; T, TNF-α; TB, TNF-α and Birinapant; TBQ, TNF-α, Birinapant, and QVD-OPh; PI, propidium iodide; h, hours; SD, standard deviation.