Figure 1.

Effects of DGKα or DGKζ deficiency on T_H differentiation. (A) Relative mRNA expression level of DGKα and DGKζ in WT CD4⁺ T cells before and after 48-h culturing in the indicated T_H and iTreg differentiation conditions. Mean ± SEM of triplicates are shown and represent three experiments. (B–H) WT, DGKα^−/−, and DGKζ^−/− naïve CD4⁺ T cells were cultured in T_H1, T_H2, and T_H17 conditions in vitro for 4–5 days. (B) Representative dot plots of cytokine-producing cells gated on CD4⁺ T cells after PMA and ionomycin stimulation for 4–5 h. (C) Bar graphs show mean ± SEM of percentages and numbers of IFNγ⁺ cells and total CD4⁺ T cells. (D) Bar graphs show mean ± SEM of percentages IL-4⁺ cells. (E) Bar graphs show mean ± SEM of IL-17A⁺ and IL-17F⁺ cells. (F) Cytokine concentrations in culture supernatants collected at 96 h. (G) Relative mRNA levels of cytokines in indicated T_H conditions after 48 h of incubation. (H) Overlaid histograms of intracellular staining of indicated transcription factors under indicated T_H conditions for 60 h. (I) Bar graphs show mean ± SEM of survival rates of CD4⁺ T cells under different T_H conditions. Data shown are representative of or pooled from at least three independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001 (Student t-test).