*2.8. Real-time Quantitative PCR and Western Blotting*

For qPCR analysis, total RNA from hippocampal tissues were extracted using trizol regent according to the manufacturer's instructions. Complementary deoxyribonucleic acid (cDNA) was obtained by using a FastQuant RT Kit and real-time PCR analysis was used with SuperReal PreMix Plus according to the 2-step reaction program. qPCR analysis was conducted using primers as follows: *gapdh* forward 5 -CATGGCCTTCCGTGTTCCTA-3 , reverse 5 -CCTGCTTCACCACCTTCTTGAT-3 , *bdnf* forward 5 -GCCTTCATGCAACCGAAGTA-3 , reverse 5 -TGAGTCTCCAGGACAGCAAA-3 .

For immunoblot analysis, hippocampal tissues were resuspended with lysis buffer. The protein was extracted and samples were resolved on 10% sodium dodecyl sulfonate–polyacrylamide gel (SDS-PAGE) and then transferred to polyvinylidene fluoride (PVDF) membranes. Antibodies against the following proteins were used: BDNF and β-tubulin goat anti-rabbit immunoglobulin G (IgG).
