Skip to main content
. Author manuscript; available in PMC: 2017 Mar 14.
Published in final edited form as: J Immunol. 2016 Jan 29;196(5):2309–2318. doi: 10.4049/jimmunol.1502074

FIGURE 5. Binding of [3H]LOS:sCD14 to V135A hMD-2 in the presence of TLR4ecd.

FIGURE 5

HEK293T cells were cotransfected with plasmids encoding hTLR4ecd and wt or mutant hMD-2, as indicated. After 24 h, the transfection medium was changed with serum-free medium and spiked with 1 nM [3H]LOS:sCD14. Medium was harvested at 24 h and analyzed by Sephacryl S500 size exclusion chromatography. Resolved reactants and products were monitored by liquid scintillation spectroscopy. Note that the reaction of [3H]LOS:sCD14 with MD-2:TLR4ecd yields ([3H]LOS:MD-2/TLR4ecd)2 complex, whereas the interaction of [3H]LOS:sCD14 with sMD-2 leads to the formation of [3H]LOS:MD-2. The results shown are representative of two or more experiments.