Supplementary Figure 4: The expression of G131D or E promotes the activation of ERK in response to weak OVA APL stimulation.
From: Slow phosphorylation of a tyrosine residue in LAT optimizes T cell ligand discrimination
a. Representative contour plot of ERK phosphorylation of G131D (left population), G131E (middle population), or WT LAT (right population) expressing J.OT-I+hCD8+ Jurkat variants as in Fig. 4a,b. Ligands used for stimulation are indicated above the plots. b. Bar graphs depicting the percent of p-ERK+ cells as shown in Fig. 4b,c. The percent of p-ERK+ cells at peptide concentration at 1000 pM represents the gated population in Fig. 4b. Each symbol represents one experiment (n = 4). Data are compiled from four independent experiments. The statistical analysis for cells stimulated with OVA peptide: ***P = 0.0001 (WT vs E at 100 pM); ***P = 0.0002 (WT vs E at 1000 pM); ****P = <0.0001. For cells stimulated with Q4R7 peptide: *P = 0.0159; **P = 0.0019; ****P = <0.0001; ns, not significant P = 0.0838. For cells stimulated with T4 peptide: *P = 0.0298; ***P = 0.001; ****P = <0.0001; ns, not significant P = 0.1385. For cells stimulated with Q4H7 peptide: ***P = 0.0009; ****P = <0.0001; ns, not significant P = 0.4285 (WT vs D at 1 pM); P = 0.8305 (WT vs E at 1 pM); P = 0.4731 (WT vs E at 10 pM); P = 0.6343 (WT vs E at 100 pM); P = 0.2393 (WT vs E at 1000 pM); For cells stimulated with G4: **P = 0.0035 (WT vs D at 10 pM); **P = 0.0048 (WT vs D at 100 pM); **P = 0.0030 (WT vs D at 1000 pM); *P = 0.0192 (WT vs E at 10 pM); *P = 0.0358 (WT vs E at 1000 pM); ns, not significant: P = 0.9978 (WT vs D at 1 pM); P = 0.2937 (WT vs E at 1 pM); P = 0.3008 (WT vs E at 100 pM). For cells stimulated with Catnb peptide: ****P = <0.0001; ns, not significant P = 0.9967 (WT vs D at 1 pM); P > 0.9999 (WT vs E at 1 pM); P = 0.6354 (WT vs E at 10 pM). For cells stimulated with VSV peptide: ns, not significant: P > 0.9999 (WT vs D at 1 pM); P = 0.8635 (WT vs D at 10 pM); P = 0.9991 (WT vs D at 100 pM); P > 0.9999 (WT vs D at 1000 pM); P = 0.9658 (WT vs E at 1 pM); P > 0.9999 (WT vs E at 10 pM); P = 0.9998 (WT vs E at 100 pM); P = 0.9688 (WT vs E at 1000 pM); One-way ANOVA test.
