Figure 4: Human sperm with reduced motility from infertile subjects exhibit a significant decrease in PI3K/AKT activity and PHB phosphorylation. The phosphorylation levels of PI3K P85 subunit α+γ (pY199 and pY467), AKT (pS473; pT308), and PHB (pT258) in human sperm from normospermic (N), asthenospermic (A) and oligoasthenospermic (OA) subjects were analyzed by western blotting. Beta-actin was used as the loading control and densitometric analysis used the relative ratios of P85 subunit α+γ (pY199 and pY467) versus P85, AKT pS473 versus AKT, AKT pT308 versus AKT, or PHB pT258 versus PHB. Fifty micrograms of protein were loaded in each lane. The detected number (n) in each cohort was indicated in each histogram. Median with interquartile range, and* P < 0.05;** P < 0.01;*** P < 0.001 determined by the Kruskal–Wallis test followed by the Dunn's posttest. PHB: prohibitin; AKT: protein kinase B; PI3K: phosphatidylinositol 3-kinase.