Ab41733

The following antibodies were used: rabbit antibodies against AdipoR2 (1:800 for WB)^{50 (link)}, TEX14 (1:1000 for WB, 1:200 for IF, Abcam; Ab41733, GR109649-1), SYCP1 (1:5000 for IF, Abcam; Ab15090, GR3184119-1), γH2AX (1:3000 for IF, Abcam; Ab11174, GR294890-8), RAD51 (1:500 for IF, Thermo Fisher Scientific; PA5-27195, UI2840658J), ELOVL2 (1:1000 for WB, Abcam; Ab176327, GR139744-4), CerS3(1:1000 for WB, 1:100 for IF)^{51 (link)}, SUN1 (1:1000 for WB, 1:100 for IF, Abcam; Ab103021, 1014380-1), KASH5 (1:1000 for IF, Hiroki Shibuya lab), MAJIN (1:1000 for IF)^{19 (link)}, TERB1 (1:1000 for IF)^{18 (link)}, and TERB2 (1:1000 for IF)^{19 (link)}; rat antibody against RPA2 (1:200 for IF, Cell Signaling Technology; 2208 S, 3); mouse antibodies against TRF1(1:1000 for IF)^{18 (link)}, MLH1 (1:50 for IF, BD Biosciences; 51-1327GR, 4136717), and β-Actin (1:2000 for WB, Sigma; A2228-200UL, 067M4856V); goat antibody against Lamin B (1:100 for IF, Santa Cruz Biotechnology; sc-6216, F1715); and chicken antibody against SYCP3 (1:5000 for IF)^{52 (link)}.

Testes were directly embedded in tissue freezing medium, sectioned at 5 µm, and fixed with 4% paraformaldehyde. Slides were blocked with bovine serum (Sigma), and stained with primary antibodies. The primary antibodies used were as follows: anti-PEX16 (1∶200; PA5-60311, Invitrogen), anti-ABCD1 (1∶200; 11159-1-AP, Proteintech, Manchester, UK), anti-ABCD3 (1∶200; PA1-650, Invitrogen), anti-TEX14 (1∶500; ab41733, Abcam, Cambridge, UK), anti-DDX4 (1∶200; ab27591, Abcam), anti-γ-H2AX (1∶200; ab26350, Abcam), anti-SOX9 (1∶200; AB5535, Millipore, Billerica, MA, USA), anti-PLZF (1∶200; AF2944, R&D Systems, Minneapolis, MN, USA). Secondary antibodies were FITC- or TRITC-conjugated goat anti-mouse or anti-rabbit IgG and donkey anti-goat IgG (1∶1000; Beijing Zhongshan Biotechnology Co., Beijing, China). The slides were visualized using a ZEISS LSM800 (ZEISS, Jena, Germany) microscope.