Sybr green pcr master one mix kit

Total RNA was extracted using TRIzol® (Thermo Fisher Scientific). PrimeScript RT kit was used to reverse the mRNA into cDNA, and the SYBR-Green PCR Master One-Mix kit (TransGen, Biotech, Co., Ltd.) was used for quantitative real-time PCR. β-actin was used as an internal control. The sense and anti-sense primers were listed in Table 1.Table 1

The primers used in this study

Gene	Sequences (5′-3′)
	Sense	Anti-sense
RBM14	CTTCGACTACCAGCAGGCTTTT	CCGTCAGAGGCGCCACATAAG
EP300	ATTAAGGAACTGGAACAGGAG	AGAGGTCGTTAGATACATTGG
HK2	TGATGTGGCTGTGGATGAGCT	GCCAGGCAGTCACTCTCAATCTG
PDK1	GTGTAGATTAGAGGGATG	AAGGAATAGTGGGTTAGG
PKM2	GCACACCGTATTCAGCTCTG	TCCAGGAATGTGTCAGCCAT
LDHA	AGGCTGGGAGTTCACCCATTAAGC	GAGTCCAATAGCCCAGGATGTG
GLUT1	TGTCGTGTCGCTGTTTGTGGTGGA	TGAAGAACAGAACCAGGAGCACAG
GLUT3	GAGGTGCTGCTCACGTCTC	TTGAATTGCGCCTGCCAAAG
β-actin	ATTGGCAATGAGCGGTTC	CGTGGATGCCACAGGACT

After 48 hours of transfection and culture, the total RNA was extracted by Trizol reagent (Invitrogen, Carlsbad,USA) for qRT-PCR detection. The SYBR Green PCR Master One-Mix kit (Transgen, Beijing, China) and cDNA Reverse Transcriptase Kit (TAKARA, Beijing, China) were purchased from ElifeBio (Hangzhou, China). Real-time PCR 7300 System (Applied Biosystems, Foster City, USA) was used for qRT-PCR detection. The reaction conditions were as follows: pre-denaturation at 95℃ for 5 min, denaturation at 95℃ for 10 s, annealing at 60℃ for 30 s, and 35 cycles. Each group was repeated 3 times. Relative quantification of RNA expression was calculated using the 2-ΔΔCT method. All the primers of HOTAIR, miR-206, CCL2 and GAPDH were shown in Table S.