Fig. 1

ScRNA-seq and microarray analysis (discovery set) showing main immune cells in the testicular microenvironment of full or dysfunctional spermatogenesis. A UMAP plots of cell clusters in all testicular samples as well as in full spermatogenesis or spermatogenic dysfunction group. Cells were colored by different cell types (left) or by different sample groups (right). B Dot plot showing expression patterns of canonical marker genes across different cell types. C Proportions of all cell types in eight testicular samples. D, E and F Immunohistochemical staining of additional markers of Mast cells (CMA1), Macrophages (CD68) and T cells (CD3Z), respectively, in the testis. Arrows represented positive cells. The original images of IHC stained sections were obtained from Human Protein Atlas database (https://www.proteinatlas.org). G Bar plot showing comparison between normal (full spermatogenesis) and disease (spermatogenic dysfunction) samples of the radio of three immune cells to somatic cells. H Bar plot showing normalized ssGSEA scores of three immune cells in different groups. Data shown as mean ± SD. *p < 0.05, **p < 0.01, ns: not significant (compared with full spermatogenesis group). I The top and bottom enriched pathway (according to NES) in GSEA of differently expressed genes compared between high versus low mast cell infiltration groups. GSEA was based on h.all.v7.4.entrez.gmt. J, K and L Scatter plots showing spearman correlations between JS and mast cells, macrophages and T cells infiltration level, respectively (reflected by ssGSEA scores, similarly hereinafter). rho, spearman correlation coefficient. Dots are colored by different JS. Dot diameter represented immune cell infiltration level. Curves were fitted with loess method. UMAP uniform manifold approximation and projection, ssGSEA single sample gene set enrichment analysis, JS Johnsen score, NES normalized enrichment score, GSEA gene set enrichment analysis