Fig. 4.

Inactivation of Runx1 in the ovarian somatic cells has minimal impacts on morphogenesis of the ovary with the exception of transcriptomes. Immunofluorescences for a pre-granulosa cell marker FOXL2, Sertoli cell marker SOX9, and germ-cell marker TRA98; b interstitial cell marker COUP-TFII (encoded by Nr2f2 gene), Laminin, and Sertoli cell marker AMH; c meiotic cell marker SYCP3 and germ-cell marker TRA98 in E14.5 XX control, XX Runx1 KO, and XY control gonads. The gray color represents DAPI nuclear staining. Single-channel images are provided in Supplementary Fig. 2. Expression of d pre-granulosa or germ-cell-specific genes and e Sertoli or Leydig cell-specific genes in XX control and XX Runx1 KO gonads at E14.5 by quantitative PCR (n = 5). Values are presented as mean ± SEM.; unpaired Student’s t-test; *p < 0.05, ***p < 0.001; NS, not significant. f Immunofluorescence for FOXL2, TRA98, and nuclear counterstain DAPI (blue) in XX control and Runx1 KO gonads at birth (P0). Scale bar: 100 µm. g Venn diagram comparing the 317 genes differentially expressed in XX Runx1 KO vs. XX Control gonads with the 749 genes differentially expressed in XX Foxl2 KO vs. XX Control gonads at birth. Forty-one percent of the genes differentially expressed in Runx1 KO (129/317) were also misregulated in the absence of Foxl2. Genes differentially expressed were identified by microarray (n = 4/genotype; fold change > 1.5, one-way ANOVA p < 0.05). h Quantitative PCR analysis of Runx1 and Foxl2 mRNA expression in XX control, Runx1 KO, and Foxl2 KO gonads at birth (n = 5/genotype). Values are presented as mean ± SEM. One-way ANOVA, p < 0.05. Bars with different letters (a, b) are significantly different. For all experiments, controls are wild-type littermates of Runx1 KO mice. Source data are provided as a Source Data file. For the immunofluorescences, at least three independent biological replicates were analyzed and the images presented are representative of all replicates