For AAV-injected mice, 2 hours after extinction learning (Ext.) or retrieval of fear memory (No Ext.), the vHPC tissues were collected and rapidly frozen in liquid nitrogen. The total RNA was extracted from the vHPC tissues using TRIzol Reagent (Invitrogen). Four micrograms of total RNA was used as template for cDNA synthesis and amplification with the FastQuant RT Kit (Tiangen) according to the manufacturer’s instructions. The cDNA was diluted to an equal concentration of 400 ng/μl, and 80 ng of which was used for further PCR amplification. Real-time PCR was then performed using the cDNA as template in a Power SYBR Green PCR Master Mix (Applied Biosystems). The primers used were as follows: Gapdh, 5′-CCTCGTCCCGTAGACAAAATGGT-3′ (forward) and 5′-TTGAGGTCAATGAAGGGGTCGT-3′ (reverse); Fos, 5′-GAGCTGGAGCCCCTGTGTAC-3′ (forward) and 5′-CAGGGTAGGTGAAGACAAAGGAA-3′ (reverse); Npas4, 5′-CAGATCAACGCCGAGATTCG-3′ (forward) and 5′-CACCCTTGCGAGTGTAGATGC-3′ (reverse); Bdnf, 5′-AGGATCCCCATCACAATCTTACA-3′ (forward) and 5′-GCCACTGACCACACAATTGCT-3′ (reverse). The plate was run in the Applied Biosystems 7500 Fast Real-Time PCR System under the Standard 7500 run mode (one cycle 50.0°C, 20 s; one cycle 95.0°C, 10 min; 35 cycles 95.0°C, 15 s and 60°C, 1 min with fluorescence measured during 60°C step). Data were then analyzed using the 2−ΔΔCt method. All collected data were normalized to the No Ext. group of AAV-Syn-GFP–injected mice.

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