Anti amh

Dissected gonads were fixed in Bouin’s solution, dehydrated, and embedded in paraffin (Paraplast, Sigma, P3683). Histological staining was performed according to Debreuill’s trichromatic (aniline blue and picric acid, Sigma, B8563) method as previously described [22 ,23 (link)]. For immunochemistry, heat-induced epitope retrieval was conducted in a sodium citrate buffer (10 mM sodium citrate, 0.05% Tween-20, pH 6) at 95 °C for 20 min as previously described [5 (link)]. Subsequently, the sections were blocked with 3% H₂O₂ and 10% goat serum (Sigma, G9023), incubated with primary antibodies (all rabbit polyclonal: anti-AMH, 1:100, Santa Cruz Biotechnology, sc-166752; anti-cleaved caspase 3, 1:100, Assay BioTech, L0104; anti-collagen I, 1:300, Abcam, ab34710; anti-Cre, 1:3000, Abcam, ab190177; anti-CYP17A1, 1:200, Abcam, ab125022; anti-N-cadherin, 1:1000, Abcam, ab76057; anti-PCNA, 1:500, Abcam, ab18197; anti-TRA98, 1:500, Abcam, ab82527) at 4 °C overnight, and stained using UltraVision Quanto Detection System (ThermoFisher, Fremont, CA, USA, TL-125-QHD). Mayer’s hematoxylin (Sigma, MHS32-1L) was used as a counterstain. Sections were viewed under the Nikon Eclipse E600 microscope. The number of animals used for histological and immunohistochemical analysis is presented in Supplementary Table S3.