Figure 3: Lineage-tracing experiments for the Wt1-positive cells in the ovary and transcriptome analysis of the mesonephros- and ovary-derived Gli1-positive cells.
(a–c) Lineage-tracing of the fetal ovary-derived Wt1-positive cells in the Wt1-CreERT2; Rosa-LSL-tdTomato; Gli1-LacZ embryos was induced by tamoxifen (TM) administration at E10.5. The ovaries were analysed at different stages of development (E11.5, P4 and 1 month) by fluorescent immunohistochemistry for tdTomato, WT1, germ cell marker PECAM-1, LacZ, granulosa cell marker FOXL2, steroidogenic marker 3βHSD and 4,6-diamidino-2-phenylindole (DAPI). E=embryonic day; P=postnatal day. d–f are higher magnification of the outlined areas in a–c. Arrows indicate fetal ovary-derived Wt1-positive cells. n=3–4 for each specimen. Scale bar, 25 μm. (g) Microarray analyses of three independent pools of the mesonephros-derived and the neonatal ovary-derived Gli1-positive cells. All cells were isolated from the adult ovaries of Gli1-CreERT2; Rosa-LSL-tdTomato mice at 2 months of age. (h) Quantitative PCR analysis of Gli1 and markers of theca cell steroidogenesis (Star, Cyp11a1, Cyp17a1 and Lhcgr) in the mesonephros-derived (n=3) and the neonatal ovary-derived Gli1-positive cells (n=3). *P<0.05; two-tailed Student’s t-test. Values are presented as means±s.e.m.
