Journal: Nature Communications

Article Title: Defective neutrophil-derived exosomes facilitate macrophage activation through miR-122-5p in Behçet’s disease

doi: 10.1038/s41467-025-63348-8

Figure Lengend Snippet: A Transmission electron microscopy (TEM) analysis of the size and shape of exosomes derived from PMN of active BD, inactive BD and HC. Arrow indicates exosome derived from PMN. B Quantity of exosomes from unstimulated (left) or stimulated (right) PMN (active BD = 23, inactive BD = 9, HC = 23). C Correlation analysis of BD PMN exosomes and CRP ( n = 32). D IL-6 and TNF production, ( E ) CD80 and ( F ) CD86 expression on HMDMs ( n = 10 for each group) stimulated with LPS and BD PMN exosomes (5ug/mL), HC PMN exosomes (5ug/mL) or PBS (blank control) for 9 h ( D ) and 4 h ( E, F ). One-way ANOVA tests ( B ) and two-way ANOVA (D-F) were used with adjusted two-tailed p-value using two-stage linear step-up procedure of Benjamini, Krieger and Yekutieli were used. Pearson correlation analysis was used in ( C ). LPS, lipopolysaccharide; MFI, mean fluorescence intensity; CRP, C-reactive protein; HMDM, human monocyte-derived macrophages. N for all experiments were biological replicates. Data were summarized as mean ± standard deviation (SD). *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001; ns, not significant.

Article Snippet: PMN (3*10 6 /ml) were stimulated by 100 ng/ml LPS (437620, Sigma) or blank control (PBS) in X-VIVO for 24 h. The supernatant was centrifuged at 2000g, 30 min to remove cells and debris, then 20 ml supernatant was mixed with 10 ml Total Exosome Isolation reagent (4478359, Life Technologies Corporation, Carlsbad, California, USA) and incubated at 4°C overnight, which was centrifuged at 10,000 g for 1 h to obtain the exosomes.

Techniques: Transmission Assay, Electron Microscopy, Derivative Assay, Expressing, Control, Two Tailed Test, Fluorescence, Standard Deviation