2.3 Western blotting
The proteins of the cultured cells and sperm samples were extracted
using a universal protein extraction lysis buffer (Bioteke) containing a
protease inhibitor cocktail (Roche). Then, denatured proteins were
separated on 10% SDS-polyacrylamide gels and transferred onto a
polyvinylidene difluoride (PVDF) membrane (Millipore) for immunoblot
analysis. The primary antibodies used were anti-SLC26A8 (1:1000, Atlas
antibody) and GAPDH (1:5000, Abcam).