Fig. 4: Apoptosis and PGCCs formation contribute to single cell derivation of GCT.

A Schematic diagram depicting the process used to conduct RNA-Seq analysis on cells from the indicated types of ovaries (n = 3 for each group). B Bar graph of the number of differentially expressed genes (DEGs) found when WT-GCs, MT-GCs, and GCTs were compared, showing that GCTs and MT-GCs shared the lowest number of DEGs among the three compared groups. C Venn diagram showing the number of DEGs in the MT-GCs vs. WT-GCs, GCTs vs. WT-GCs, and GCTs vs. MT-GCs comparisons, revealing that 1,760 DEGs were up/downregulated in GCTs compared to those in WT-GCs and MT-GCs. D Significantly enriched pathways of the up-regulated differentially expressed genes (DEGs) between GCTs and the other two groups. Data were analyzed by DAVID Bioinformatics Resources 6.8 for Gene Ontology (GO) enrichment analysis of DEGs. GO terms with corrected P-values that lower than 0.05 were considered significantly enriched by DEGs. E Heat map of apoptotic genes in WT-GCs, MT-GCs and GCTs groups, showing upregulation of apoptosis related genes during tumorigenesis. F TUNEL detection in ovaries of No-Cre control and DKO mice, demonstrating the upregulation of apoptosis signals during cyst formation. Representative images are shown. Scale bar: 100 μm. G Statistical results of F. Data represent mean ± SD. *P < 0.05, unpaired t test. H Hematoxylin staining and fluorescent detection of ovaries at different developmental stage of Rb-DKO ovaries, showing PGCCs (arrowheads) formation was involved in the tumor formation. Boxed regions in upper panels are magnified in the lower panels. Scale bar: 10 μm. I Heat map of genes responses to hypoxia among WT-GCs, MT-GCs and GCTs, indicating a high level of hypoxia response in GCTs. J Heat map of stemness genes among WT-GCs, MT-GCs and GCTs, illustrating cancer stem cells were involved in tumor formation and progression.