Single-cell analysis of Treg patients with acute graft-versus-host disease. (A) Single cells from the indicated CD4⁺ T-cell populations from aGVHD patients were sorted into 96 cells plates without prior stimulation and analyzed using 48.48 Dynamic Arrays. The heat map of the single-cell gene expression profiling was obtained by 2-dimensional hierarchical clustering using Euclidean distance and average linkage. (B) Single-cell mRNA expression of Treg markers (FOXP3, IL2RA, CTLA4, Helios, LSGALS, and IKZF4), chemokine receptor CCR8 and the housekeeping gene B2M were plotted for RA⁻DR⁻ and RA⁻DR⁺ Tregs and Tconvs. One of 3 analyses of 3 different patients is shown.