FIGURE 6.

EA treatment restored downregulation of neuroprotective p-GSK-3β, p-Akt, BDNF, and Bax in the WD-TBI mouse cortex. (A,C) Representative Western blot results show levels of p-GSK-3β, p-Akt (A), and mature BDNF (C) in the mouse cortex from the Control, WD, and WD+EA groups. β-Actin was used as the loading control. (B,D) Quantitative analysis of p-GSK-3β, p-Akt, and mature BDNF protein expression levels in (A,C), respectively. (E) The relative mRNA expression levels of BDNF in the cortex 48 h after TBI were evaluated by qPCR. (F) Representative images of IHC staining show anti-BDNF antibody immunoreactivity in cortex samples in the Control, WD, WD+EA, and WD+EA-NAP groups. High magnification images marked by the small red boxes from the top panels (green box) are shown below (red box). The bottom panels in (F) show higher-magnification images of the cortex from the middle panels marked by the small black boxes. Scale bars of the green box panels are 250 μm; of the red box panels are 100 μm. (G) Representative Western blot results show levels of Bax expression in cortex from the Control, WD, and WD+EA groups 48 h after TBI. β-Actin was used as the loading control. (H) Quantitative analysis of Bax protein expression levels in (G). Data are the means ± S.E.M. of at least three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 (statistical testing was performed by one-way ANOVA/Newman-Keuls test). The number underneath each bar in (B), (D), (E), and (H) refers to the number of mice used in the study group. The original Western blot images are presented in Supplementary Figures 7–9. The F values of one-way ANOVA for (B), (D), (E), and (H) are presented in Supplementary Table 5.