*4.10. Statistical Analysis*

The continuous variables were expressed as mean ± standard error of the mean (SEM). The statistical method was used to compare all the continuous variables with one-way analysis of variance (one-way ANOVA). Once the ANOVA was significant, for post hoc assessment, we used Scheffé multiple range test to assess the difference between groups, especially the experimental group and the control group. A *p* < 0.05 was considered statistically significant.

#### **5. Conclusions**

In the present study, our results suggested activation of VEGF/VEGFR2 (Flk-1) by PGC-1α subsequently activates PI3K/AKT and MEK/ERK signaling by alternating the activation state of numerous of downstream proteins relevant to cell proliferation and inhibiting apoptosis in the hippocampus following status epilepticus. Otherwise, downregulation of PGC-1α by siRNA against to *pgc-1*α will inhibit VEGF/VEGFR2 (Flk-1) signaling and suppress pro-survival PI3K/AKT and MEK/ERK pathways that also are accompanied by hippocampal CA3 neuronal cell apoptosis. These results may indicate that the PGC-1α induced VEGF/VEGFR2 pathway may trigger neuronal survival signaling and PI3K/AKT and MEK/ERK signaling pathways. Thus, the axis of PGC-1α/VEGF/VEGFR2 (Flk-1) and downstream PI3K/AKT and MEK/ERK signaling could be considered as an endogenous neuroprotective effect against apoptosis in the hippocampus following status epilepticus. Taking together, our previous research and the results from this study, new strategies for treatment, disease modification, and neuroprotection in epilepsy, particularly in status epilepticus, should be developed.

**Author Contributions:** Conceptualization, Y.-C.C., H.-Y.P., S.-P.H., and S.-D.C.; methodology, T.-K.L., X.-P.L., C.-W.L., and J.-H.L.; investigation, J.-B.H., S.-F.C., and Y.-C.C.; data curation, X.-P.L., J.-H.L., T.-K.L., C.-W.L., and N.-C.C.; writing—original draft preparation, J.-B.H., S.-P.H., and Y.-C.C.; writing—review and editing, Y.-C.C. and S.-D.C.; visualization, J.-B.H., S.-F.C., C.-Y.H., and H.-Y.P.; supervision, Y.-C.C.; project administration, Y.-C.C.; funding acquisition, Y.-C.C. and H.-Y.P.; statistical analysis, H.-Y.C. All authors have read and agreed to the published version of the manuscript.

**Funding:** This work was supported by research grants MOST102-2314-B-182A-090-MY3 from the Ministry of Science and Technology, Taiwan to Yao-Chung Chuang, and CMRPG8I0041 from the Chang Gung Medical Foundation to Hsiu-Yung Pan.

**Conflicts of Interest:** The authors declare no conflict of interest.
