RT-qPCR Analysis of lncRNA and mRNA

The cells were first dissolved in 1 mL TRIzol (Thermo Fisher Scientific), and the total RNA was extracted according to the manufacturer's specifications. The total RNA was reverse transcribed into cDNA using M-MLV reverse transcriptase (D7160L, Beyotime) and random primers. The reaction system was configured according to the Premix EX Taq™ II Kit (Takara, Dalian, China), following the manufacturer's instructions. An ABI7500 quantitative PCR (Applied Biosystems, Shanghai, China) was used for RT-qPCR with glyceraldehyde phosphate dehydrogenase (GAPDH) as a loading control for lncRNA and mRNA. The relative expression level was determined by the 2^−ΔΔCt method [22 (link)]. The primers used are listed in Table 1.