Fig. 1

Single-cell RNA-seq analysis of Lin28⁺ spermatogonia isolated from adult testes. (A) Whole-mount images of subsets of Lin28-YFP⁺ cells in seminiferous tubules from Lin28-YFP transgenic mice at 2 months. (B) t-SNE plot of Lin28-YFP-positive spermatogonial clusters defined by scRNA-seq analysis. Each dot represents a single cell, and clusters are marked by different colors. (C) Gene expression patterns of selected marker genes projected on t-SNE plots. (D) Violin plots of SSC, progenitor and differentiated spermatogonial marker expression in different clusters. (E) Monocle pseudotime trajectory analysis of Lin28-YFP-positive spermatogonial subsets defined in clusters. Cells are marked by pseudotime scores, with dark colors representing immature stages and light colors representing mature stages. (F) Heatmaps of pseudotime-dependent genes and enriched KEGG terms for each temporal stage. Blue indicates low expression, and red indicates high expression. (G) Volcano plot of genes differentially expressed between putative SSCs in the C8 cluster and other clusters (adjusted p value < 0.05; fold change > 2). Red indicates upregulated genes, and blue indicates downregulated genes