Skip to main content
. 2020 Jun 24;11:1122. doi: 10.3389/fimmu.2020.01122

Figure 3.

Figure 3

High expression of disulphide-HMGB1 in human leukocytes. (A) Western blot probed with anti-CD45, anti-HMGB1, and anti-GAPDH antibodies in reducing conditions (upper panels) or probed with anti-HMGB1 antibody in non-reducing conditions (lower panel) on peripheral blood mononuclear cells (PBMCs) isolated from four healthy human donors. The upper band corresponds to the fully reduced-HMGB1 (frHMGB1) and the lower band to the disulphide-HMGB1 (dsHMGB1) in the lower panel. (B) Quantification of HMGB1 redox isoforms expression normalized on Ponceau staining. A.U. = arbitrary unit (n = 4 healthy donors). (C,D) Western blot probed with anti-HMGB1 antibody in non-reducing conditions on PBMCs stimulated with anti-CD3/anti-CD28 antibodies or lipopolysaccharide (LPS) for 24 or 72 h (C). Percentage of HMGB1 redox isoforms expression (D). Ctrl, control unstimulated cells (n = 2 healthy donors). Data represent the means ± SEM and statistical significance was calculated by Two-way ANOVA (D).