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. 2018 Dec 21;9:3070. doi: 10.3389/fimmu.2018.03070

Figure 6.

Figure 6

TLR9 but not RIG-I activation requires glycolysis to induce the production of type I IFNs in primary human pDCs. Freshly isolated primary human pDCs were stimulated with 1 μM of CpG-A in the absence or presence of 2-deoxy-D-glucose (2-DG; 5 mM) then IFN-α and IFN-β protein levels (A,B) and lactate concentrations (C) were measured from the supernatants of the cells at 12 h. In parallel experiments cells were pre-treated with low dose of CpG-A for 16 h then following thorough washing steps stimulated with 5′ppp-dsRNA (RIGL, 1 μg/ml) alone or in combination with 5 mM of 2-DG or left untreated. IFN-α and IFN-β protein levels (F,G) and lactate concentrations (H) were measured from the supernatants at 6 h. (D,E,I,J) Cell viability was measured by 7-aminoactinomycin D (7-AAD) staining using flow cytometry. (D,I) Representative dot plots are shown where numbers indicate the percentage of 7-AAD negative cells. (A–C,E,F–H,J) Data represent the mean ± SD of three individual experiments. *p < 0.05, ***p < 0.001, ****p < 0.0001 vs. control; #p < 0.05, ##p < 0.01, ###p < 0.001, ND, not determined.