Fig. 4

Differential Effects of PMT Inhibition on Murine and Human Th1 Cell Differentiation. a, b CD4⁺ T cells from the spleen and peripheral lymph nodes of IFN-γ-YFP reporter mice were enriched and polarized under Th0 (IL-2) or Th1 cell conditions in the absence (Th0) or presence of indicated probes (1 μM; red) or their controls (where available; black). a Flow cytometric analysis of intracellular YFP reporter signal (representing IFN-γ expression) was detected at day 4. b Secreted IFN-γ was analyzed by ELISA in the supernatant of the same experiment. Each data point represents one of three biological replicates and the data shown is representative of three independent experiments. c, d CD4⁺ T cells from the blood of three healthy human donors were cultured under Th0 or Th1 cell-polarizing conditions in the presence or absence of indicated probes or their controls. c Flow cytometric analysis of intracellular IFN-γ was detected at day 4. d Secreted IFN-γ was analyzed by ELISA in the supernatant of the same experiment. Each data point represents one of three donors. Dotted lines visualize the mean frequency of IFN-γ-positive Th1 cells in the absence of probes (a, c) or the mean concentration of IFN-γ in the supernatant (b, d). Significant differences are indicated with an asterisk and were calculated using one-way ANOVA (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001). e Western blot analysis of the effect of indicated inhibitors (red) or control compounds (black) on the trimethylation of H3K27 and dimethylation of H3K79 in CD4⁺ T cells under Th1, Th2, Th17, Treg cell-polarizing conditions. Please see Supplementary Figs. 5a-5d for information on the MW markers. Data shown is representative of 2 independent experiments. Error bars represent SEM. See also Supplementary Figs. 2-5