Supplementary Figure 7: T cell ligand discrimination is preferentially regulated at LAT Y132.

a. CD8-negative or positive LAT-deficient J.OT-I⁺ Jurkat variants were reconstituted with wild-type LAT or G131D LAT (J.OT-I⁺.hCD8neg.LAT.WT, J.OT-I⁺.hCD8neg.LAT.G131D, J.OT-I⁺.hCD8⁺.LAT.WT, or J.OT-I⁺.hCD8⁺.LAT.G131D). Cells were stimulated with T2-K^b antigen-presenting cells pulsed with OVA peptide, OVA APL peptides, self-peptide Catnb, or VSV control peptide over a wide range of peptide concentrations as in Fig. 3. The percentage of cells that are CD69⁺ is plotted against peptide concentration (mean ± s.d; n = 3 technical replicates). Data are representative of three experiments. b. Statistical analysis of CD69 upregulation as in (a) with the stimulation of 10 nM of each peptide (mean; n = 3 technical replicates). Data are representative of three experiments. c. LogEC50 analysis of CD69 induction assays as in (a). d. LAT-deficient J.OT-I⁺ hCD8⁺ Jurkat variants were reconstituted with wild-type LAT or D126G-Y127, D170G-Y171, E190G-Y191, or D225G-Y226 LAT. Cells were subjected to CD69 induction assays as in (a). The percentage of cells that are CD69⁺ is plotted against peptide concentration (mean ± s.d; n = 3 technical replicates). Data are representative of two experiments.