Fig. 4.

PDIA3 is detected in greater abundance in spermatozoa derived from the low quality fractions. A, Human sperm ejaculates were taken from individual donors and high- and low-quality fractions were isolated through Percoll density gradients. Nuclei isolated from spermatozoa were then lysed and separated via SDS-PAGE. Following transfer, the nitrocellulose was probed with primary antibodies against PDIA3. Cross-reacting bands were visualized with chemiluminescence after addition of horse-radish peroxidase conjugated secondary antibody. The approximate position of the molecular mass markers are shown. B, An equal sized aliquot of the same sample used for immunoblot was loaded into SDS page, run and total protein was detected via silver stain. C) PDIA3 abundance according to the SWATH analysis between high- and low-quality sperm cells demonstrates significantly more abundance within the latter (n = 8, *p < 0.01).