Figure 2.

Kinetics of the suppressive effect of DMXAA in THP1 dual reporter cells. (A) PMA-differentiated THP1 dual reporter cells were stimulated with various concentrations of DMXAA (1, 3, 10, 30, or 100 µg/ml) together with increasing concentrations of c-di-AMP (3, 10, or 30 µg/ml) for 24 h in three independent experiments. IRF activity was determined by measuring secreted luciferase activity in the supernatants. Representative data from three independent experiments are shown as the mean ± SD of triplicates (**p < 0.01, ****p < 0.0001, One-way ANOVA with Sidak’s multiple comparison test). (B) IFNβ production was measured using ELISA. Representative data from three independent experiments are shown as the mean ± SD of triplicates (**p < 0.01, ***p < 0.001****, p < 0.0001, One-way ANOVA with Sidak’s multiple comparison test). (C) PMA-differentiated THP1 dual reporter cells were stimulated with DMXAA (100 µg/ml) at the indicated time points (−3, 0, or 8 h) together with c-di-AMP (30 µg/ml) for 24 h in three independent experiments. IRF activity was determined by measuring secreted luciferase activity in the supernatants. IFNβ production was measured using ELISA. Representative data from three independent experiments are shown as the mean ± SD of triplicates (*p < 0.05, **p < 0.01, Student’s t test). (D) NF-κB activity was determined by measuring secreted embryonic alkaline phosphatase (SEAP) activity in the supernatant. Representative data from three independent experiments are shown as the mean ± SD of triplicates (*p < 0.05, **p < 0.01, ***p < 0.01, Student’s t test).