Quantscript rt kit

Total RNA was extracted from NP cells using Trizol reagent (TransGen Biotech) according to the manufacturer's instructions. Total RNA (1 μg) was reverse‐transcribed into cDNA by using the Quantscript RT Kit (Beyotime) according to the manufacturer's protocol. The cDNA was then used as the template for the qPCR with TransStart® Top Green qPCR SuperMix kit (TransGen Biotech) on the Eppendorf Realplex4 instrument (Eppendorf). The reaction conditions were set as follows: 95°C for 3 min, followed by 40 cycles at 95°C for 30 s and at 55°C for 20 s, and extension for 20 s at 72°C. All primer sequences are listed in the Table 1. The 2^−ΔΔCt method was used to calculate the relative RNA expression levels of target genes.³⁴

Total RNA was extracted from cells using Trizol reagent (TransGen Biotech, Beijing, China) according to the manufacturer’s protocols. cDNA was synthesized from 1 μg of total RNA by using the Quantscript RT Kit (Beyotime, Shanghai, China) according to the supplier’s protocol. cDNA was amplified using a TransStart^® Top Green qPCR SuperMix kit (TransGen Biotech, Beijing, China) on the Eppendorf Realplex4 instrument (Eppendorf, Hamburg, Germany). The reaction conditions were as follows: 95 °C for 3 min, followed by 40 cycles at 95 °C for 30 s and at 55 °C for 20 s, and extension for 20 s at 72 °C. The sequences of primers were presented in Table 1. The 2^−ΔΔCt method was used to calculate the relative RNA expression levels of target genes.