Total RNA was extracted from tissues and cells using TRIzol reagent (Takara, Beijing, China). The complementary DNAs (cDNAs) for the LncRNAs and mRNAs of interest were reverse-transcribed from 2 μg total RNA using PrimeScript RT-polymerase (Takara). The cDNAs for the miRNAs of interest were synthesized from 1 μg total RNA using miRNA 1st Strand cDNA Synthesis SuperMix (Vazyme, Nanjing, China). We performed qRT-PCR using SYBR-Green Premix (Takara), miRNA Universal SYBR® qPCR Master Mix (Vazyme), and specific PCR primers (Sangon Biotech, Shanghai, China). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and U6 were used as internal controls. We determined the expression of LncRNA, mRNA, and miRNA using the calculating 2−ΔΔCT method. Primer sequences are summarized in Table .
Primer Sequences
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