Fig 4. Temporal changes in the expression levels of pre-granulosa or Sertoli cell-specific genes in ovarian grafts in the male-host environment.

(A) Hierarchical clustering of microarray data obtained from the grafted ovaries on days 0, 4, 7, 10, 15, and 20 post-transplantation, based on all 45,101 probes in the Mouse Genome 430 2.0 GeneChip array. Duplicate raw data were generated from transplants on days 0, 10, and 20 post-transplantation. (B) GO terms (p<0.01) for 241 down/576 upregulated genes (|fold| > 2, false discovery rate [q] < 0.3) in grafted ovaries on day 20 post-transplantation. (C) Venn diagrams showing 643 pre-granulosa (orange) and 695 Sertoli cell (blue)-specific gene probes selected from among the altered gene probes at each stage by comparing data from Sertoli cells versus pre-granulosa cells at 12.5 dpc using a published dataset [47] (right circles). (D) Scatterplot showing the number of down/upregulated pre-granulosa/Sertoli cell-specific genes in the grafted ovaries. Each point on the graph shows the grafted ovaries on each day post-transplantation. The vertical or horizontal lines show the total numbers of downregulated pre-granulosa or upregulated Sertoli cell-specific genes (|fold| > 2, p<0.05) at 12.5 dpc from the published dataset [42]. (E) Heatmap showing relative changes in expression levels of the 525 pre-granulosa (containing cluster I) and 402 Sertoli cell-specific genes (containing clusters II/III) in grafted ovaries (signal intensity: high [red], medium [black] to low [green]). (F) GO terms (p<0.05) for cluster I and clusters II/III genes in the ovarian grafts. (G, H) Line graphs show the fold expression changes for several key pre-granulosa (G) and Sertoli cell-specific genes (H) post-transplantation. The expression levels on day 0 post-transplantation were set as 1.0 on the y-axis.