Fig. 6.

Acidic amino acid residues in the juxtamembrane region of TLR5 are important for UNC93B1 binding and signaling. (A) TLR5 was immunoprecipitated from RAW cells stably expressing UNC93B1-HA with TLR5-GFP (WT or DE/ST). Coimmunoprecipitated UNC93B1-HA (Top) as well as UNC93B1-HA (Middle) and TLR5-GFP (Bottom) in the input lysate were detected by immunoblotting. The data shown are representative of three independent experiments. (B) BMDCs expressing WT or DE/ST mutant TLR5-GFP were imaged. (C) BMDCs expressing GFP or TLR5-GFP (WT or DE/ST mutant) were stimulated with flagellin (100 ng/mL) or LPS (100 ng/mL) for 4 h and the intracellular TNF-α levels in the GFP-positive cells were measured (open histograms). Data shown are representative of three independent experiments. Shaded histograms represent the levels of TNF-α in unstimulated cells.