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. 2021 Jan 8;11:581728. doi: 10.3389/fimmu.2020.581728

Figure 1.

Figure 1

The change in T cell subtypes induced by combination therapy with metformin and tacrolimus. (A) The proportion of Th1 and Th17 cells in normal mice treated with combination therapy. CD4+T cell isolated from normal C57BL/6 mice were stimulated with anti-CD3 (0.5 ug/ml) in the presence of Metformin or FK506 alone or combined Metformin and FK506 for 3 days and analyzed by flow cytometry. A plot from one representative experiment displays the proportions of IL-17+, IFN-γ + among CD4+ T cells. (B) The difference in proportions of Th1 and Th17 cells in mice with graft-versus-host disease after combination therapy. CD4+T cell isolated from GVHD mice were stimulated with anti-CD3 (0.5 ug/ml) in the presence of Metformin or FK506 alone or combined Metformin and FK506 for 3 days and analyzed by flow cytometry. A plot from one representative experiment displays the proportions of IL-17+, IFN-γ + among CD4+ T cells. Numbers in the plots indicate percentages of gated cells. Data are means ± SEMs. Data are representative of three independent experiments. (*p < 0.05, **p < 0.01, ***p < 0.005) (C) Gene expression analysis by RNA sequencing before and after treatment with combination therapy. CD4+T cell isolated from normal C57BL/6 mice were stimulated with anti-CD3 (0.5 ug/ml) in the presence of Metformin or FK506 alone or combined Metformin and FK506 for 3 days and analyzed by RNA-seq data. Heatmaps of genes encoding for molecules involved in Th1, Th17 cells, and Treg cell function that are differentially expressed in combination of metformin and FK506 compared to vehicle.