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. 2014 Nov 3;9(11):e111460. doi: 10.1371/journal.pone.0111460

Figure 1. Transient transfection of TLR4 Asp299Gly, TLR4 Thr399Ile or TLR4 Asp299Gly/Thr399Ile with MD2 into HEK cells results in constitutive NF-κB activation but a reduced relative increase compared to basal NF-κB activity.

Figure 1

HEK cells were transiently transfected with wild-type or mutant TLR4 +MD-2 +CD14, together with the reporter constructs NF-κB-luc and Renilla luciferase. After 48 h the cells were stimulated for 6 h with medium alone (C) or medium +10 ng/ml LPS (LPS) and the luciferase activity was determined in cell lysates. Data are from a representative experiment (n = 3 experiments) and are shown as mean ±SEM for that experiment. A) Relative luciferase activity of wild type TLR4, Asp299Gly TLR4, Thr399Ile TLR4 or Asp299Gly/Thr399Ile TLR4 with or without stimulation with 10 ng/ml LPS. B) Fold increase in luciferase activity compared to basal activity induced by stimulation of wild type TLR4, Asp299Gly TLR4, Thr399Ile TLR4 or Asp299Gly/Thr399Ile TLR4 with 10 ng/ml LPS. C MD2  =  unstimulated cells in the presence of MD2; LPS MD2  =  LPS stimulated cells in the presence of MD2; C-MD2  =  unstimulated cells with MD2 absent; LPS-MD2  =  LPS stimulated cells with MD2 absent.