Fig. 5: Effects of aging on granulosa cell functions.

A UMAP plot of granulosa cell (GC) subcluster analysis from combined single cell datasets is shown. B Changes in the proportion of the GC subclusters with age are shown, as determined using permutation testing. Error bars indicate the 95 percent confidence interval for the magnitude of the difference between ages as determined by bootstrapping. Dashed lines indicate absolute log₂-fold changes greater than 0.58, and false discovery rate less than 0.05. Gray points indicate no significant difference between ages. C Spatial distribution of cell subtypes within a single late antral-stage follicle (highlighted in navy, top left panel). Bar plot (top right) shows the fraction of each subtype within the selected follicle determined by cell type transfer from the reference dataset. The location of each subtype (bottom left), as well as the distribution of distances for each cell to the center of the follicle (bottom right) is highlighted for each subtype. D Bar plot highlights the relative signaling for all pathways identified as significantly altered in GCs (>20% change in strength, p-value <= 0.01) between young and old datasets. E Heatmaps show the relative contribution of each GC subtype for each identified differential signaling pathway, shown for incoming and outgoing signaling in both age groups. Colors represent relative contribution for each cell type (values scaled row-wise with row max = 1). Dendrograms represent the results of hierarchical clustering of both rows and columns. F Bar plots show the identity of cell types receiving signaling from GCs, split by age. G Bar plots show the identity of cell types sending signals to GCs, split by age. H Bar plots depict the strength of total signaling from GC subtypes to myeloid cells, split by age. I Bar plot highlighst the strength of total signaling sent from late antral/mural GCs to myeloid subtypes, split by age. J Chord plots show SEMA3 (top) and Desmosterol (bottom) signaling from GC subtypes to myeloid subtypes. Chord plot signaling is depicted by lines connecting subtypes, and the strength of signaling is depicted through the thickness of lines in the young (left) and old (right) datasets. Major source cell types are bold/black, and target cell types are bold/green.