1,25(OH)2D3 induces pERK1/2, AKT and GSK3 phosphorylations in human sperm through VDR. A: Percoll-purified sperm washed spermatozoa were incubated in the unsupplemented Earle's medium for 30 min at 37°C and 5% CO2, in the absence (NC) or in the presence of increasing 1,25(OH)2D3 concentrations (0.01 nM, 0.1 nM, 1 nM and 10 nM) or with 0.1 nM 1,25(OH)2D3 combined with anti-VDR Ab (AbVDR). Other samples were incubated in capacitating medium (Cap). B: Time course study (0, 5, 10, 30 and 60 min) of ERK1/2, AKT and GSK3 phosphorylations treated with 0.1 nM 1,25(OH)2D3. Actin was used as loading control. On the side are reported the densitometric evaluations normalised against actin levels. The autoradiographs presented are representative examples of experiments that were performed at least seven times with repetitive results. *P < 0.05 versus control, **P < 0.01, ***P < 0.002 versus control.