FIGURE 4.

Knockdown of DAI blocks ALD-DNA-induced macrophage activation in vitro. A and B, RAW264.7 cells were transfected with a plasmid vector encoding siControl or siDAI, stimulated with ALD-DNA (4 μg/ml), and then subjected to real-time PCR (A) and Western blot analysis (B) to evaluate the expression of DAI. Data in B are representative of results obtained in three independent experiments. C and D, RAW264.7 cells (C) and BMDMs (D) were transfected with siControl or siDAI, and treated with PBS, UnALD-DNA (4 μg/ml), or ALD-DNA (4 μg/ml) for 24 h. Levels of TNF-α, IL-6, IL-12, IL-1β, IL-10, MCP-1, IFN-β, and nitrite (the inducible NO synthase, iNOS) in the culture supernatants of macrophages were measured by ELISA. Data are mean ± S.E. of three independent experiments. *, p < 0.05; **, p < 0.01; ***, p < 0.001; NS, not significant.