qRT- PCR
Total RNA from hippocampus and NSCs was extracted using Trizol reagent
and following the manufacturer’s instructions. The quality of RNA was
determined by the ratio of A260/A280,and samples with
A260/A280=1.8~2.0 were used for further
analysis.Followed the instructions of the cDNA Reverse Transcription Kit
to reversed transcribe RNA into cDNA. Fluorescence quantification was
performed using the TBGreen® Premix Ex Taq™ II (Takara, Japan), and then
the amplifying conditions for cDNA were as follows: 95°C for 30s, 40
cycles of 95°C for 5s and 60 °C for 30s. Primer3 software
(http://simgene.com/Primer3) was used to design specific primers. Primer
sequences are listed in Table 1.