Intrathecal(i.t.) injection of ephrinB1-Fc induced a dose- and time-dependent increase of spinal PI3K-p110γ and p-AKT expression.

<p>The expression of spinal PI3K-p110γ and p-AKT was assayed at 0, 0.17-h, 0.5-h, 1-h, 4-h, and 8-h time-points in the time-dependent experiments and at 0.5-h time-point in the dose-dependent experiments after ephrinB1-Fc injection. PI3K inhibitor wormannin (0.4 µg) or LY294002 (5 µg) or DMSO was intrathecally injected at 30 min before i.t. ephrinB1-Fc (0.5 µg),the expression of p-AKT in spinal was assayed at 1 h after ephrinB1-Fc injection in PI3K mediated spinal AKT activation experiment. (A, B) The representative bands (top) for the expression of PI3K-p110γ or p-AKT in spinal at different time points after injection of ephrinB1-Fc (0.5 µg) and the quantitative data (bottom) for the expression of PI3K-p110γ or p-AKT. The fold change for the density of PI3K-p110γ or p-AKT normalized to GAPDH or AKT for each sample, respectively. The fold change of PI3K-p110γ or p-AKT level in 0 time point groups was set at 1 for quantifications. *<i>P</i><0.05, **<i>P</i><0.01, compared with 0 time point group, n = 4 mice in each group. (C, D) The representative bands (top) for the expression of PI3K-p110γ or p-AKT in spinal after injection of various dose of ephrinB1-Fc (0.02, 0.1, and 0.5 µg). The quantitative data (bottom) for the expression of PI3K-p110γ or p-AKT. The fold change for the density of PI3K-p110γ or p-AKT normalized to GAPDH or AKT for each sample, respectively. The fold change of PI3K-p110γ or p-AKT level in Fc groups was set at 1 for quantifications. *<i>P</i><0.05, **<i>P</i><0.01, compared with Fc group, n = 4 mice in each group.(E) The representative bands (top) and the quantitative data (bottom) for the expression of spinal p-AKT. The fold change for the density of p-AKT normalized to AKT for each sample. The fold change of p-AKT level in DMSO-saline group was set at 1 for quantifications.*<i>P</i><0.05, compared with DMSO-ephrinB1-Fc group, n = 4 mice in each group.