Fig. 6. Evaluation of cAMP levels and tyrosine phosphorylation status following WGA treatment. To investigate the production of intracellular cAMP following WGA challenge, human spermatozoa were incubated with either WGA (0.5 µg/ml) or SJA (0.5 µg/ml), or in positive (+ve) or negative (–ve) control media for 30 minutes. Following incubation, the sperm sample was split and one half was prepared for (A) the measurement of [cAMP]_i and the remaining cells were prepared for (B) immunodetection of phosphotyrosine residues (anti-py). This experiment was replicated three times with pooled semen samples obtained from at least three different donors and representative blots are presented. Graphical data represent the means ± s.e.m. from the three replicate experiments. **P<0.01.