Fig. 2
Molecular signatures of SSCs containing undifferentiated spermatogonial subpopulations. (A) t-SNE plot of the reclustering of putative SSCs containing spermatogonia. Each dot represents a single cell, and cell clusters are distinguished by color. (B) t-SNE plot with cell cycle analysis of putative spermatogonial stem cells. (C) Relative expression of markers for SSCs, progenitors and differentiating spermatogonia. (D) Pseudotime trajectory analysis of the origin subsets of undifferentiated spermatogonia defined in clusters. Cells are colored according to the pseudotime score, with dark colors representing immature stages and light colors representing mature stages. (E) Heatmaps of pseudotime-dependent genes and enriched KEGG terms for each temporal stage. Blue indicates low expression, and red indicates high expression. (F) Detection of FOXF1, FOXC2, FOXP1 and EOMES with undifferentiated spermatogonia markers (LIN28A and ZBTB16) by immunofluorescent whole mount staining. Examples of As, Apr, and Aal (up to 16 interconnected cells) spermatogonia are shown by white dotted lines. White arrows indicate negative expression. Scale bars = 100 μm, n = 3