Fig. 2

HMGB1 diminished the immunosuppressive effect of MSCs in vitro. MSCs were first treated with or without IFN-γ and TNF-α (5 ng/ml) in the presence or absence of HMGB1 (1 μg/ml) for 24 h, then co-cultured with mice spleen-derived CD4⁺ T cells at ratios of 1:20, 1:40 and 1:80 (MSCs: T cells) for 72 h. a T cell proliferation was measured by CFSE assay. b The percentage of IFN-γ-positive T cells (gated on CD4⁺ cells) was assessed by flow cytometry to represent the T cell activation. Column scatter graphs were used to visualize the comparison. Each data point represented the average value of triplicated wells from each independent experiment. Data were pooled from at least three independent experiments. Data were shown as mean ± SD. *P < 0.05