FIG 6.

MyD88 and IRAK-1 are involved in VpOmpU-mediated proinflammatory responses. (A) Increase in gene expression of MyD88 in VpOmpU-treated RAW 264.7 and THP-1 cells at different time points. Fold change is calculated as value above that for buffer-treated cells, and results are expressed as mean ± SD from three independent experiments. (B) MyD88 coimmunoprecipitated with TLR2 upon VpOmpU treatment in RAW 264.7 cells as evidenced by immunoblotting and densitometric analysis of the bands. (C) MyD88 coimmunoprecipitated with TLR2 upon VpOmpU treatment in THP-1 cells as evidenced by immunoblotting and densitometric analysis of bands. Results are expressed as mean ± SEM from three independent experiments (*, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, P > 0.05 [versus band intensities in the buffer-treated cells]). (D) Decrease in TNF-α and IL-6 production in VpOmpU-treated BMDMs differentiated from MyD88^−/− mice compared to the wild-type control. Results are expressed as mean ± SEM from three independent experiments (*, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, P > 0.05 [versus VpOmpU-treated BMDMs from wild-type mice]). (E) Decrease in TNF-α production with use of IRAK-1 inhibitor in response to VpOmpU treatment in RAW 264.7 and THP-1 cells. Results are expressed as mean ± SEM from three independent experiments (*, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, P > 0.05 [versus cells treated with VpOmpU only]).