FIGURE 7.

Inhibition of NADPH oxidase suppresses LPS or lauric acid-induced TLR4 dimerization. A, Ba/F3 cells were pretreated with DPI (2 μm) for 1 h or NAc (10 mm) for 2 h and then stimulated with LPS (100 ng/ml) or (B) 100 μm lauric acid for 30 min. Cells were lysed, and GFP-TLR4 was immunoprecipitated and immunoblotted with anti-FLAG and anti-GFP antibodies. C, RAW264.7 cells were pretreated with 2 μm DPI for 1 h and treated with 100 μm lauric acid for 30 min. Cells were lysed, and lysates were fractionated by sucrose gradient fractionation. Lipid raft fractions (1–3) were collected, trichloroacetic acid-precipitated, and immunoblotted with anti-TLR4 and anti-flotillin-1 antibodies.