Impaired CARD11-JNK signaling leads to increased NFAT2 expression and activation. (A) Volcano plot showing DEGs from the comparison of WT Jurkat T cells stimulated with 20 ng/ml PMA and 1 μg/ml ionomycin (P/I) for 2 h after a 2-h pre-treatment with JNKi versus those stimulated with P/I for 2 h after pre-treatment with DMSO. The upregulation of NFATC1 (NFAT2) is highlighted, and the expanded plot shows normalized counts for NFATC1 (NFAT2) across experimental conditions. (B) Quantitation of the fold change in NFATc1 (NFAT2) DNA binding, normalized to unstimulated WT (NS), in WT or CARD11 KO Jurkat T cells stimulated with P/I for 4 h (n = 3–4). (C) Quantitation of the fold change in NFATc1 (NFAT2) DNA binding, normalized to unstimulated WT (NS), in WT Jurkat stably transduced with WT or L194P and stimulated with P/I for 4 h (n = 3). (D) Expression of GATA3 and NFATC1 (NFAT2) in CADINS patient versus HD naïve CD4 T cells after 3 days of anti-CD3/anti-CD28 activation. (E) Schematic summarizing the hypothesized effects of WT or DI CARD11 on NFAT2 and GATA3 expression and activity. Error bars indicate the SEM. Asterisks denote significance by two-way ANOVA with Šídák’s multiple comparisons test. Created in BioRender. Bauman, B. (2025) https://biorender.com/a16y201. *P < 0.05; **P < 0.01; ***P < 0.001; and ****P < 0.0001. NS/No stim, no stimulus.