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. 2015 Jul 6;112(29):E3883–E3892. doi: 10.1073/pnas.1501662112

Fig. S2.

Fig. S2.

EZH2 differential expression is not efficient enough to enhance CD8+ T-cell effector function. (A and C) Representative flow cytometric dot plots showed that, compared with transfection with siRNA-Ctrl or medium, siRNA-EZH2 did not induce significant changes of percentages of IFN-γ– or TNF-α–producing CD8+ T cells. Data were gated on CD8+ T cells. PBMCs from patients with active TB were transfected with siRNA targeting EZH2 (siRNA-EZH2) or siRNA-Ctrl or transfection medium for 48 h and cultured for another 3 d. Cells were then harvested and analyzed for the expression of EZH2 in CD8+ T cells and intracellular IFN-γ and TNF-α responses of CD8+ T cells using ICS/flow cytometry. (B and D) Pooled data show the percentages of IFN-γ– or TNF-α–producing CD8+ T cells in response to knock down of indicated siRNA or medium (n = 10). (E) Representative CBA assay of PBMC of a patient with active TB showing that, compared with transfection with siRNA-Ctrl or medium, siRNA-EZH2 did not induce significant changes of concentration of TNF-α and IFN-γ. The red and green squares mark the TNF-α and IFN-γ, respectively. (F and G) Pooled data showing the concentrations of IFN-γ and TNF-α upon transfection of indicated siRNA or medium (n = 10). (H) ChIP-qPCR analysis of H3K27Me3, EZH2, and H3Ac at the promoters of IFN-γ and TNF-α in CD8+ T cells transfected with siRNA-EZH2 and siRNA-Ctrl. NS, no statistical significance. Error bars represent SEM from two independent experiments.