Skip to main content
. Author manuscript; available in PMC: 2015 Sep 1.
Published in final edited form as: J Immunol. 2013 Dec 11;192(2):714–721. doi: 10.4049/jimmunol.1302384

FIGURE 1. Mutagenesis of the 2DL4 cytoplasmic domain identifies critical residues for signaling.

FIGURE 1

A, The 2DL4 cytoplasmic tail truncations, as indicated, were transfected into HEK293T cells. After 48 h, IL-8 secretion was measured by ELISA. IL-8 secreted after transfection of wt 2DL4 was 430 pg/ml. The data shown is representative of 2 to 4 experiments for each truncation. B, The TRAF6-binding motif P-X-E-X-X-Ar/Ac, (Ar, aromatic; Ac, acidic) in the 2DL4 cytoplasmic tail is aligned with similar motifs in human CD40, IRAK, and TRANCE-R. The aromatic Y277 in the 2DL4 cytoplasmic tail was mutated to either aromatic F or aliphatic A. IL-8 secretion was measured 48 h after transfection of HEK293T cells with 2DL4 and the Y277F or Y277A mutants. IL-8 secreted after transfection of wt 2DL4 was 1750 pg/ml. The data shown is representative of 2 experiments. C, Plasmids encoding point mutations of the indicated 2DL4 cytoplasmic tail residues were transfected into HEK293T cells. After 48 h, IL-8 secretion was measured by ELISA. Mock Tf: Mock-transfected cells. Results represent the mean ± SEM of triplicate samples from one representative experiment out of 3 experiments. IL-8 secreted after transfection of wt 2DL4 ranged from 888 to 1110 pg/ml.