Fig. 6: Interactome analysis of PI16⁺ RCs during homeostasis and inflammation.

a, Gene sets enriched upon inflammation based on differentially expressed genes in tonsillitis (t)PI16⁺ RCs compared to PI16⁺ RCs of patients with OSA. Numbers highlight top genes with an average log₂ (fold change) (avg. log₂ (FC)) > 1 assigned to enriched terms. b, Violin plots show gene expression profiles in different PI16⁺ RC clusters. c, Scatterplot indicates interaction strengths of individual cell subsets based on all signaling pathways derived. Dot size reflects interaction count. d, Hierarchical plots show inferred interactions of collagen and FN1 signaling networks. e, Circle plots represent inferred TGF-β and FGF signaling networks. d,e, Edge width reflects the communication probability. f, Significantly different (paired Wilcoxon test) relative information flow of signaling pathways between adult patients with OSA and tonsillitis. g–i, Bulk tonsillar fibroblasts from n = 8 (medium, TGF-β1, LIGHT), n = 6 (FGF2, TGF-β1 + LIGHT) and n = 5 (TGF-β3, FGF2 + LIGHT) patients with OSA (g) or sorted TRCs and PI16⁺ RCs from n = 5 adult patients with OSA or tonsillitis (h,i) were stimulated with recombinant proteins for 48 h. PI16 (g) and THBS1 (i) mRNA fold change measured by quantitative PCR with reverse transcription (RT–qPCR). h, IL-6 concentration in culture supernatants. j–m, Carboxyfluorescein succinimidyl ester (CFSE)-labeled T cells were cultured (3 d) with PI16⁺ RCs and recombinant proteins and were stimulated with PHA. j, Representative flow cytometric plots depicting CD25 expression on CFSE^loCD4⁺ T cells. k, Quantification of CFSE^lo cells as a percentage of CD4⁺ cells. l, Quantification of CD25⁺ cells as a percentage of CFSE^loCD4⁺ cells. m, IL-10 concentration in culture supernatant. g–i, Box extends from 25th to 75th percentiles, median is indicated and whiskers show the minimum and maximum values. k–m, Mean and s.d. are indicated, n = 4 patients with OSA or tonsillitis. q values were calculated with the Kruskal–Wallis (g,i), Friedman (h) or repeated measure (RM) one-way ANOVA (k–m) test and Benjamini, Krieger and Yekutieli correction for multiple comparisons. Interactome analysis represents scRNA-seq data of 81,997 T and B cells from n = 2 adult patients with OSA and n = 3 adult patients with tonsillitis and 23,704 FRCs from n = 5 adult patients with OSA and n = 4 adult patients with tonsillitis. CTL, cytotoxic T lymphocytes; T_CM, central memory T cells; T_reg, regulatory T cells; T_FH, follicular helper T cells; MBC, memory B cells.