Single-cell RNA sequencing reveals the altered transcriptomic profile of patients with dysfunctional NFATc1. Basal transcriptome and profiled TCR repertoire of lymphocytes from P1 and P2 compared with that of HCs via single-cell RNA sequencing (5′ scRNAseq, 10x Genomics). (A) Analysis of TCR antibody repertoire diversity with a tree-map representation for patients P1 and P2 and age-matched HCs. Each square represents a unique clone and its size reflects its productive frequency within the repertoire. (B) Uniform manifold approximation and projection plot of the combined scRNAseq data set from P1, P2, and HCs. (Left) Cells colored based on cell type annotation after mapping to a reference data set of healthy PBMC,¹⁹ cell types with <20 cells omitted for clarity. (Right) Cells colored based on the genotype; patient data shown above HC data. (C) Bar graphs showing proportions of different cell populations within the lymphocytes. (D) Heatmap showing pseudobulk expression data for genes (shown as rows) that are differentially expressed between patients and HCs within CD4⁺ and CD8⁺ T-cell populations (samples and cell types shown as columns). Genes shown are the top 20 up- or downregulated in any of the 2 tests (tests performed with edgeR software with exact test and pseudobulk data; false discovery rate < 0.05; ranked based on the P value). Expression values shown are scaled log counts per million cropped to the range −2 to 2. (E) Heatmap showing single-cell expression data for genes differentially expressed between patients and HCs within CD8⁺ T-cell subpopulations (naïve, TCM [central memory T cells] and TEM [effector memory T cells]). Genes shown are the top 30 up- or downregulated in any of the 2 tests (tests performed as in panel D). (F) Heatmap showing single-cell pseudobulk expression data for genes differentially expressed between patients and HCs within CD19⁺ B cells (tests performed as in panel D).