Fig. 3: Comparative analysis of regenerative and homoeostatic Aundiff by single-cell RNA-seq.
From: Distinctive molecular features of regenerative stem cells in the damaged male germline
a Volcano plot of DEGs (MAST differential expression test with Bonferroni correction, adjusted P value < 0.05) within Gfra1 + cells (normalised expression level > 2) from scRNA-seq analysis of sorted Aundiff (E-Cadherin + α6-integrin+ c-KIT–) from CTRL and BU-treated mice at D10 (755 Gfra1 + cells in CTRL and 1331 in BU). Genes of interest are highlighted. b GO of DEGs from comparison of Eomes + (345 cells in CTRL and 667 in BU) or Gfra1 + cells from scRNA-seq analysis of a. P values from one-sided Fisher’s Exact test. c UMAP plots showing clustering analysis of scRNA-Seq data of CTRL and D10 BU Aundiff of a (3798 cells in CTRL and 2669 in BU-treated). Expression of selected genes associated with Aundiff (left panels), and genes upregulated in regenerative Aundiff (right panels) are shown. d Representative wholemount IF of tubules D10 post-BU (n = 3 per group). Arrows: uPAR+ As cells. Scale bar: 50 μm. Dashed lines indicate seminiferous tubule profiles. e Representative flow cytometry analysis of adult mice treated with BU vs. CTRL at D10. Graph shows mean percentage of Aundiff (E-Cadherin+ c-KIT–) uPAR+ ± SEM (n = 5 per group). Significance determined by two-tailed unpaired student t test. f Venn diagrams illustrating overlap of up and downregulated DEGs (Fold change > 1.5 and adjusted P value < 0.05) within Gfra1 + spermatogonia from CTRL vs. BU-treated Aundiff from analysis of a and CTRL (adult) vs. neonatal ID4bright spermatogonia36. P values derived from hypergeometric tests are shown. Examples of concordantly regulated DEGs are indicated. Source data are provided as a Source Data file.
