Ab26350

Human seminiferous tubules were fixed with 4% paraformaldehyde and stored in 70% ethanol. Then, fixed samples were embedded in paraffin and sliced into 5-μm sections by microtome (Leica RM2235). Immunostaining was performed as described previously (42 (link)). Deparaffined and rehydrated sections were washed in PBS for 5 min after antigen retrieval and then blocked with 5% BSA at room temperature for 1 h. Subsequently, the sections were incubated with primary antibodies at 4°C for more than 8 hours and secondary antibodies (Jackson ImmunoResearch, Alexa 488-, Alexa 594-) for 1 h at room temperature. Nuclei were counterstained with 10 μg/ml Hoechst 33342 for 15 min at room temperature. All images were captured with a ZEISS LSM880 confocal microscope. The experiments performed in this study were approved by the Third Affiliated Hospital of Guangzhou Medical University (2017-055). And informed consents to every donor in our study have been signed.
The following primary antibodies were used: mouse anti-γH2AX (Abcam, ab26350), mouse anti-FGFR3 (Santa Cruz, sc-13121), mouse anti-BMPR1B (Abcam, ab1565836), mouse anti-DDX4 (Abcam, ab27591), rabbit anti-INHBB (Abcam, ab69286), mouse anti-ACVR2B (R&D Systems, MAB3392), rabbit anti-pSMAD2 (Cell Signaling Technology, #18338), rabbit anti-GDF5 (Abcam, ab93855), rabbit anti-pSMAD1 (Abcam, ab226821) and rabbit anti-ACVR1B (Abcam, ab109300).