Figure 3. LAG3 association with the TCR-CD3 complex increases LAG3 proximity to the co-receptors.

(a) Dynamic association of LAG3, TCR and CD4 on Lag3^+/+ CD4⁺ T cells stimulated with TCRβ Ab using sensitized emission FRET with quantification shown (Scale bar = 5μm). (b) Representative super resolution STORM of molecular distribution of CD4, LAG3 and TCR within the IS determined in CD4⁺ T cells isolated from spleen and lymph nodes of Lag3^+/+ mice and stimulated with TCRβ Abs. Three-dimensional intermolecular analyses of STORM molecule data and the median shortest distance from LAG3 to TCR and CD4 (top) or TCR to LAG3 and CD4 (bottom) (scale bar = 1μm). (c) Representative confocal image of phase separation assay using lipid bilayers containing TAMRA labeled CD4^CT or CD8^CT in the presence or absence of LAG3^CT, with LAG3^CT labeled with AF647 to visualize as a control (scale bar = 1μm). Quantification of phase separation assay with standard deviation as a measure of homogeneity. Data in (a, b and c) represent analyses from at least 3–4 independent experiments and statistics determined by unpaired Student’s two-sided t test or Wilcoxon matched pairs signed rank test with P values noted in figure.