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. 2015 Apr 13;112(17):5455–5460. doi: 10.1073/pnas.1422576112

Fig. 3.

Fig. 3.

Pocket residues serve divergent roles in TLR2/1 and TLR2/6 responsiveness. (A and B) HEK293T cells were transiently transfected with reporter constructs for endothelial leukocyte adhesion molecule (ELAM)-luciferase; Renilla-luciferase; and pcDNA3.1, WT pcDNA3-YFP-hTLR2, or mutant TLR2 constructs in the same vector. Cells were pretreated for 1 h with media, vehicle (65 μM NaOH), or C29 (50 μM) and treated with P3C or P2C (50 ng/mL) for 5 h in the presence of media, vehicle, or C29. Lysates were prepared, and the dual-luciferase assay was performed. (C and D) HEK293T cells were transiently transfected with pcDNA3.1, WT pcDNA3-YFP-hTLR2, or mutant TLR2 constructs in the same vector. Western blot analysis was performed using whole-cell lysates (WCL) (C) or membrane extracts (D) to ensure comparable expression of each TLR2 mutant. Pan-Cadherin was probed as a loading protein for membrane extracts. A and B represent the mean ± SEM from two independent experiments, each carried out in duplicate, and C and D are representative of two independent experiments.