Fig. 10. Investigation of the role of sperm PECAM-1 in WGA induced signalling. To investigate the role of sperm PECAM-1 as a functional receptor for WGA signalling in human spermatozoa we examined whether anti-PECAM-1 antibodies could suppress the response to WGA. For this purpose, human spermatozoa were incubated with either anti-PECAM-1 antibodies alone or with anti-PECAM-1 antibodies followed by WGA. Control treatments in which sperm were incubated with normal rabbit serum were also included. Each incubation was conducted for a period of 30 minutes. After incubation solubilized sperm proteins were prepared for immunodetection of phosphotyrosine residues (anti-py). Blots were then stripped and reprobed with anti--tubulin to ensure equivalent protein loading across all treatments. The experiment was replicated three times with pooled semen samples obtained from at least three different donors and representative blots are depicted.