Fig. 4

Pseudotime trajectory analysis delineated the temporal dynamics of GCs during ovarian aging. A Pseudotime heatmap showing dynamic gene expression profiles during GC fate commitment. The four gene sets were determined by k-means clustering according to their expression patterns. The expression level of dynamic genes from high to low is indicated by a colour gradient from red to blue. B The top 5 enriched GO terms for each gene set are shown based on the dynamic genes of GCs. The corresponding clusters' GO terms are represented in the same colour. C Visualization of expression trends of the signature genes over pseudotime in GCs. D Expression of AMH in GCs. The colour indicates the level of expression. E Immunostaining of follicles for AMH. Three different ovaries were immunostained in each age group. Scale bars are 20 μm. F Scatterplot showing the differential trajectories of three GCs subtypes over pseudotime by Monocle. Arrows indicate that the mural GC developed into atretic and antral GCs. State refers to the fact that during the development and differentiation of GCs, different genomes are expressed (some genes are activated, while others are silenced). Branch points refer to the changes in gene expression/the emergence of new cell types during the process of cell development and differentiation