Effects of STO-609 on the CaMKIV phosphorylation in chicken sperm.

Sperm incubated at 35°C in the –5mM Ca²⁺ or +5mM Ca²⁺ medium with or without CaMKKs inhibitor STO-609 (0; 1; 10; 50 μM) at 5 min and 15 min. The control (Ctrl): sperm incubated with or without Ca²⁺ in the absence of STO-609. Proteins from sperm lysates were analyzed by western-blotting using anti-phospho-Thr196-CaMKIV or anti-CaMKIV as the primary antibody. Bands for phospho-Thr196-CaMKIV were detected at 60kDa (top lanes). Total CaMKIV (60kDa) was used as loading control (bottom lanes) and the phosphorylated protein CaMKIV (Thr196)/total CaMKIV ratio is shown at the bottom. Results express the mean ± SEM of the mean from 4 different experiments. There were no significant differences between treatments (P<0.05).