Dr5000 uv vis spectrophotometer

We extracted the total RNA with the Trizol reagent (Thermo Fisher Scientific, Inc. Waltham, MA, USA) and measured its purity and concentration with a DR5000 UV-Vis spectrophotometer (BioRad, Hercules, CA, USA). We used the TaqMan miRNA reverse transcription kit (Thermo Fisher Scientific, Inc. Waltham, MA, USA) to perform the reverse transcription of RNA into cDNA. Reverse transcription and PCR amplification and quantification of RNA were performed on SYBR Premix Ex Taq™ (Takara, Shiga, Japan) and the PCR instrument produced by Applied Biosystems (item number: 7500). The design and synthesis of primers were completed by Sangon Biotech (Shanghai) Co., Ltd. BACE1: sense primer, 5′-CACTCTGTTCTGGGTGGTCC-3′; antisense primer, 5′-CATGGGGGATGCTTACCAGG-3′. Internal reference (GAPDH): sense primer, 5′-CGGATTTGGTCGTATTGGG-3′; antisense primer, 5′-CTGGAAGATGGTGATGGGATT-3′. MiR-29c-3p: sense primer, 5′-GAAGCACCATTTGAAATCAG-3′ and antisense primer, 5′-TTGGCACTAGCACATT-3′. Internal reference (U6): sense primer, 5′-CTTCACGAATTTGCGTGTCAT and antisense primer, 5′-GCTTCGGCAGCACATATAC-3′. PCR conditions: predenaturation at 95°C for 10 min, then denaturation at 95°C for 15 s, and annealing and extension at 60°C for 60 s. The results were calculated using 2^−∆∆ct.