Figure 3.

Metformin inhibits T cell activation and p-S6 in vitro. Mice were injected i.v. with 1.8 × 10⁹ pfu of AdLacZ and sacrificed at 7 dpi. Lymphocytes from liver and spleen were isolated and cultured with the indicated concentrations of metformin (0, 5 and 10 mM) for 6 and 12 h. Cells were stimulated with PMA and ionomycin in the presence of GolgiStop during the last 5 h, followed by intracellular staining of IFN-γ, TNF-α, IL-2 and p-S6. (A) Percentages of IFN-γ⁺ TNF-α⁺ T cells from livers. (B) Percentages of IFN-γ⁺ IL-2⁺ T cells from livers. (C) Percentages and mean fluorescence intensity (MFI) of p-S6⁺ in liver and spleen T cells. Data are representative of at least three independent experiments. Values are shown as mean ± SEM of n = 3-4 samples/group from single experiments representative of at least three experiments performed. Two-tailed unpaired T test was used for statistical analysis. *p < 0.05; **p < 0.01; ***p < 0.001.