Prime script reverse transcript kit

The brain was dissected from 8-week-old mice. Total RNA was extracted from the brain tissues using TRIzol reagent (Invitrogen, #15596026) according to the manufacturer’s instructions. The integrity and quality of the isolated RNA were determined by the Nanodrop and the Bioanalyzer. The Prime script reverse transcript kit (Takara, #RR037A) was used to synthesize the cDNA. The RT-PCR experiments were conducted using the GoTaq G2 Hot Start Master mix (Promega #M7422). The PCR primers were listed in the S5 Fig. The RT-PCR were conducted using the following setting: 94°C for 3 minutes, followed by 32–40 cycles of thermocycling (94°C for 30 seconds, 52°C-56°C for 30 seconds, 72°C for 30 seconds), and 72°C for 3 minutes.

Total cellular RNA was extracted using the RNeasy Mini Kit (Qiagen, Venlo, The Netherlands). The total RNA was reverse transcribed to complementary DNA using a PrimeScript™ reverse transcript kit obtained from Takara Bio (Shiga, Japan). The expression of p53, bax, and gapdh was quantified using TaqMan Gene Expression Analyzer (Thermo Fisher Scientific). Real-time PCR was performed using a QuantStudio^® 3 PCR system (Applied Biosystems Real-time PCR systems, Thermo Fisher Scientific).