Fig. 2. Receptor molecules implicated in LPS-cellular activation present in lipid rafts after LPS stimulation. MonoMac-6 cells were stimulated with 10 ng/ml LPS in 5% HPS for 30 minutes prior to solubilisation with 1% Triton X-100 buffer for 1 hour on ice and then subjected to sucrose density gradient centrifugation. Fractions were collected from the top of the gradient, 1% n-octylglucoside was added to each fraction, and equivalent portions of each fraction were analysed by SDS-PAGE and immunoblotting. The lipid raft marker was detected using HRP-conjugated cholera toxin (A), the nitrocellulose membranes were also probed with 26ic (CD14-specific mAbs) (B), hsp70 (C), hsp90 (D), CXCR4 mAbs (E) and GDF5 polyclonal serum (F), as well as with the HTA125 TLR4-specific mAbs (G). The relative positions of the raft and non-raft (soluble) fractions are indicated.