Fig. 4. Single-cell transcriptomics of subclustered lymphocytes.

a, b t-SNE plot of the 26 lymphocyte clusters (a) and detailed lineages in different tissue contexts (b). c t-SNE plots of canonical marker genes for 26 lymphocyte clusters. d Lymphocyte proportions for LES tissue and blood between achalasia and controls, colored by cell types. e Venn diagram showing the core signature shared by T_RM and tissue-resident NK and B cells. f Pseudotemporal transcriptional trajectory of the T cells using the DPT and Slingshot algorithms. g, h Multicolor IHC and quantifications of CD4⁺ T_RM (g) and CD8⁺ T_RM (h) in the LES tissue between achalasia (CD4⁺ T_RM, n = 7; CD8⁺ T_RM, n = 8) and controls (n = 5). The T_RM were increasingly infiltrated and surrounding the residual sparse myenteric plexus (shown by PGP9.5) in achalasia. Scale bar, 20 μm. Data are represented as mean ± SD. Statistics: two-tailed unpaired t-test. *P < 0.05; **P < 0.01. Arrow, CD4⁺ and CD8⁺ T_RM. Source data are provided as a Source Data file.