2.4. Quantitative reverse transcription polymerase chain reaction (qRT-PCR)

RNA was isolated from tissues and cells using Trizol reagent (Solarbio, Beijing, China). For RNase R digestion analysis, RNA (2 μg) was treated with or without RNase R (Seebio, Shanghai, China) for 30 min. Subsequently, the complementary DNA (cDNA) was synthesized using specific reverse transcription kits (Takara, Dalian, China). Then, RNA levels were detected using SYBR Premix Ex Taq (Takara) and calculated using the 2−ΔΔCt method. The PCR amplification procedure included 95°C for 10 min, followed by 40 cycles of 95°C for 5 s, 60°C for 10 s, and 72°C for 10 s. β-Actin or U6 was considered as an internal control. The primers are presented in Table 2.

The primer sequences for qRT-PCR

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