NFAT1-deficient patient CD4⁺T cells are exhausted. (A) Volcano plot showing DE in CD4⁺ memory T cells between patient and 5 healthy controls. (B) Violin plots of significantly differentially expressed genes of interest in CD4⁺ memory T cells in patient and healthy controls. (C) Frequency of CD4⁺ T cell subsets in the patient and a representative control, including CD45RA⁺CD27⁻ TEMRA, CD45RA⁺CD27⁺ naïve (marked as “N”), CD45RA⁻CD27⁻ effector memory (EM), and CD45RA⁻CD27⁺ central memory (CM). Quadrants corresponding to each subset are shown at top right. (D) Quantification of C. (E) Expression of NFAT1 in patient and control naïve and memory CD4⁺ T cells. Mean fluorescence intensities are indicated. (F) Frequency of T_H1 (IFN-γ⁺IL-17A⁻), T_H17 (IL-17A⁺IFN-γ⁺) cells in memory CD4⁺ T cells of the patient and a representative control. (G-H) Quantification of F. (I) Frequency of T_H2 (IL-4⁺IL-21⁻) and IL21⁺ (IL-21⁺IL-4⁻) cells in memory CD4⁺ T cells of the patient and a representative control. (J-K) Quantification of I. (L) Frequency of TNF-α⁺ and IL-2⁺ CD4⁺ memory T cells in the patient and a representative control. (M-N) Quantification of L. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, ∗∗∗∗P < .0001, one-way analysis of variance and Tukey’s post-hoc test. NS, not significant. Green circles, healthy control; blue circle, II-2; blue upright triangle, I-2; blue inverted triangle, I-1; blue square, II-3; red diamond, II-1.