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. 2017 Jun 28;114(29):E5891–E5899. doi: 10.1073/pnas.1701990114

Fig. S1.

Fig. S1.

Membrane binding properties of CD3 signaling proteins in native TCR−CD3 complex. Membrane binding properties of cytoplasmic domains of CD3ε, CD3δ, and CD3γ chains in native TCR−CD3 complex were measured by dequenching FRET. The FRET donor was the mTFP fused to the C terminus of CD3 cytoplasmic domain, and the FRET acceptor was R18 dye inserted into the plasma membrane. (A) The FRET efficiency. The (B) surface mTFP level and (C) surface R18 staining level and (D) photobleaching efficiency of 293FT cells with different CD3 constructs were comparable. Each dot represents the value from one cell; n = 35, 28, and 31 for hCD3εCD-mTFP, hCD3δCD-mTFP, and hCD3γCD-mTFP, respectively. The result was a representative of five independent experiments. (Scale bar, 7.5 μm.) Error bars represent mean ± SD. P values were determined by the one-way ANOVA with Tukey's multiple comparisons test; n.s., not significant; ****P < 0.0001. (E) Representative pictures of each CD3 construct.