CADINS patient T cells exhibit decreased NF-κB and JNK signaling and increased GATA3 accumulation. (A) Top: Schematic of CARD11 protein structure with the approximate location of each patient variant denoted with a black arrow and the variant of interest. Bottom: Schematic of resulting CARD11 protein structure expressed from patient allele with splice site variant (c.220+1G>A). (B) Schematic of experimental timeline for the activation and expansion of healthy control and patient T cells from thawed PBMCs. (C) Representative immunoblot analysis of JNK and NF-κB signaling in healthy control and patient T cells stimulated for 30 or 60 min with 20 ng/ml PMA and 1 μg/ml ionomycin (P/I). Each patient was assayed with one to two healthy controls. (D) Quantitation of the expression of each protein relative to healthy control levels at the specified time point from immunoblots in C. Patient variants are color coded by protein domain location. (E) Representative immunoblot analysis of GATA3 accumulation in healthy control and patient T cells stimulated for 2 h with P/I in the presence or absence of 80 ng/ml anisomycin. (F) Quantitation of the fold change of GATA3 (left) and c-Jun (right) with P/I stimulation alone (circles) or P/I plus anisomycin (squares) relative to each respective no stimulus condition from the immunoblots in E. Patient variants are color coded by protein domain location. Error bars indicate the SEM. Asterisks denote significance by (D) one-way ANOVA with Dunnett’s multiple comparisons test and (F) two-way ANOVA with Šídák’s multiple comparisons test. *P < 0.05; **P < 0.01; ***P < 0.001; and ****P < 0.0001. CC, coiled-coil; CD, CARD; and L, LATCH. Source data are available for this figure: SourceData F5.