FIGURE 5.

Reconstitution with locked MIF trimers increases IL-8 synthesis by HPCEC. A, native PAGE analysis of E. coli-derived rMIF N110C and rMIF. The N110C rMIF and rMIF monomers were separated on native PAGE and visualized by Western blotting analysis using anti-MIF IgG recognizing both MIF monomers and trimers. B, HPCEC were treated with MIF-specific siRNA to reduce MIF levels, infected with P. aeruginosa strain 6294 at multiplicity of infection 30, and reconstituted with either 2 or 0.2 μm of purified recombinant MIF N110C trimers or rMIF monomers. Tissue culture supernatants were collected at 6 h after infection and analyzed for IL-8 levels by ELISA. Data are presented as mean values ± S.D. and are representative of two independent experiments.