Fig. 8

Overexpression of exosomal HMGB3 enhances the release of proinflammatory cytokines and the migration of macrophages. (A-B). RT–PCR and western blot analysis of the transfection efficiency of the pcDNA3.1-HMGB3 plasmid in RAW264.7 cells. (C). Western blot analysis of HMGB3 expression in exosomes derived from macrophages transfected with pcDNA3.1-vector or pcDNA3.1-HMGB3. (D). ELISA analysis of the levels of IL-1β, IL-6 and TNF-α in the SN of RAW264.7 macrophages treated with PBS, Vector-Exo, or HMGB3-Exo. n = 3 each group. (E-F). Transwell assay of the migration of RAW264.7 macrophages treated with PBS, Vector-Exo, or HMGB3-Exo. The scale bar represents 50 μm. (G-H). Western blot analysis of the expression of p-p65, p-STAT3, p-ERK1/2, p-p38, CCR2 and pro IL-1β in RAW264.7 macrophages treated with PBS, Vector-Exo, or HMGB3-Exo. # indicates that the data were compared between the Vector-Exo group and the HMGB3-Exo group. The data are representative of three individual experiments and expressed as the mean ± SEM. The data were analysed by Student’s t test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns = not significant. Abbreviations SN = cell culture supernatant; Vector-Exo = exosomes derived from macrophages transfected with the vector plasmid; HMGB3-Exo = exosomes derived from macrophages transfected with the HMGB3 plasmid