Immunohistochemistry and Immunofluorescence of Mouse Testes

Mice testes for IHC and immunofluorescence staining were proceeded as described previously.^{33 (link), 34 (link)} The experiments were repeated three times. The antibodies used for IHC/immunofluorescence staining are listed in Supplementary Information. Specifically, antigen retrieval was carried out by heating slides in a pressure cooker (121 °C) for 10 min in a citrate buffer pH9.0 (S2367, DAKO, Glostrup, Denmark) for p53 staining and pH6.0 (S2369, DAKO) for other antibody staining. The peroxidase-conjugated secondary antibodies used were mouse/rabbit EnVisionC (DAKO) for HRP-immunostaining or anti-rabbit/mouse Alexa 488/568 IgG (Invitrogen, Carlsbad, CA, USA) for immunofluorescence staining. Images were captured with a Zeiss AxioImager upright microscope and Olympus FV1000 upright confocal microscope.^{34 (link)}