Mirna 1st strand cdna synthesis supermix

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 1.Table 1

Primer Sequences

Gene name	Primer Sequence
MONC	Forward: CCAGACTTGTCGCACGGAT
	Reverse: AATGCACAGCAATCAGTTCCTC
miR-636	Forward: TGTGCTTGCTCGTCCCG
	Reverse: AGTGCAGGGTCCGAGGTATT
	RT Primer: GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACTGCGGG
GLCE	Forward: TGAACCACGTGGCCAAACAA
	Reverse: TTCGTTCCCCTCTCGTCTCC
GAPDH	Forward: GCACCGTCAAGGCTGAGAAC
	Reverse: TGGTGAAGACGCCAGTGGA
U6	Forward: AGAGAAGATTAGCATGGCCCCTG
	Reverse: ATCCAGTGCAGGGTCCGAGG
	RT Primer: GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACAAAATA

Total RNA was extracted from tissues or cells using TRIzol (Takara, Dalian, China). LncRNAs and mRNAs were reverse transcribed to the complementary DNA (cDNA) using a PrimeScript RT-polymerase (Takara, Dalian, China). The cDNAs from the miRNAs were synthesized with miRNA 1st Strand cDNA Synthesis SuperMix (Vazyme, Nanjing, China). SYBR Green Premix (Takara) on a 7500 real-time PCR system (Applied Biosystems) was used to perform quantitative real-time PCR (qRT-PCR) or miRNA Universal SYBR^® qPCR Master Mix (Vazyme) with specific PCR primers (Sangon Bio-tech Co., Ltd, Shanghai, China). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and RNU6 (U6) were selected as internal references. Relative quantification (2 − ΔΔCt) method was used for fold-change calculation. The primer sequences are listed in Supplementary Table S2.