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. 2020 Jul 16;117(5):1358–1371. doi: 10.1093/cvr/cvaa207

Figure 1.

Figure 1

High salt activates and promotes immunogenic IsoLG-adduct formation in human monocytes independently of osmolarity. Human monocytes were isolated by magnetic separation and cultured for 48 h in normal salt media (150 mMol/L NaCl), high-salt media (190 mMol/L NaCl), or normal salt media with added mannitol (80 mMol/L) as an osmotic control for 48 h. (A) Gating strategy to identify monocytes. (B) Representative and group flow cytometric data for monocyte intracellular staining of IsoLG-protein adducts (n = 16). (C) Representative histogram and group data for surface expression of CD83 on monocytes (n = 12). (D) Representative histogram and group data for monocyte surface expression of CD86 (n = 16). (E) Production of cytokines IL-6 (n = 18), (F) TNF-α (n = 18), and (G) IL-1β (n = 18) determined by flow cytometry using bead-based immunoassay on media of monocytes exposed to normal salt, mannitol, and high salt. Monocytes from each individual were exposed to three experimental conditions. The data point joined lines represent paired analyses for each human subject. Friedman nonparametric analysis with a Dunn’s post hoc test was employed in panels B to G. Clinical data of cell donors are shown in Supplementary material online, Table S5.