Sybr green premix ex taq 2 kit

Total RNA of hepatic cell lines was extracted with TRIzol reagent (15596-026, Invitrogen) and reverse transcribed to cDNA using Prime RT reagent kit (RR047A, Takara). qPCR analysis was performed with SYBR Green Premix Ex Taq II kit (Takara, RR820A) and ABI ViiA^TM 7 Real-Time PCR System. The sequences of primers used were as followings: ACLY 5′-GAC​TTC​GGC​AGA​GAC​AGG​TAG-3′ (sense) and 5′-TCA​GGA​GTG​ACC​CGA​GCA​TA-3′ (antisense), FASN 5′-GGA​TCA​CAG​GGA​CAA​CCT​GG-3′ (sense) and 5′-GCT​GTG​GTC​CCA​CTT​GAT​GA-3′ (antisense), ACC 5′-GCC​TCT​CAG​CTG​GTC​AGA​TTC-3′ (sense), and 5′-CTG​GTT​CAG​CTC​CAG​AGG​TT-3′ (antisense), SCD1 5′-AGC​AGG​TAA​ATT​GTC​GGG​GG-3′ (sense), and 5′-ACT​TTT​TAC​CCC​GAG​CCA​GG-3′ (antisense), ATGL 5′-TGA​GAG​GGG​AGG​TTT​CCA​CA-3′ (sense), and 5′-CAG​CAG​GCC​ATG​AAA​AAC​GG-3′ (antisense), HSL 5′-AAC​CCA​AGA​GGA​AGT​GCC​AT-3′ (sense), and 5′-GCT​CTA​GCG​GGG​TTA​TAG​GC-3′ (antisense).

Total RNA from cells was extracted by using the TRIzol reagent (Invitrogen) according to the manufacturer's protocol. For the quantification of miR‐629, cDNA was synthesized by the One Step Prime script miRNA cDNA synthesis kit (Qiagen, Valencia), and real‐time PCR was performed using the miRNA‐specific TaqMan MiRNA Assay Kit (Applied Biosystems, Foster City, USA) on an ABI7900 system (Applied Biosystems). For the mRNA detection, mRNA was reversely transcribed into cDNA using PrimeScript RT reagent kit (Takara, Dalian, China), and real‐time PCR was performed using SYBR Green Premix Ex Taq II kit (Takara) on an ABI7900 system (Applied Biosystems). U6 and GAPDH were used as internal controls for miRNA and mRNA expression, respectively, and the relative expression of respective genes was calculated by 2^−ΔΔCt method.

Total RNA was extracted using the RNA-Quick Purification Kit (Shanghai Yishan Biotechnology Co., Ltd, Shanghai, China) and converted into cDNA using PrimeScript RT Master Mix (TaKaRa, Dalian, Liaoning, China). Quantitative RT–PCR was carried out using the SYBR Green Premix Ex Taq II kit (TaKaRa). The primers used were as follows:
human IL-6 forward: 5′-ACTCACCTCTTCAGAACGAATTG-3′;
human IL-6 reverse: 5′-CCATCTTTGGAAGGTTCAGGTTG-3′;
human CCL2 forward: 5′-CCATGGACCACCTGGACAAGCA-3′;
human CCL2 reverse: 5′-GGTGTCTGGGGAAAGCTAGGGG-3′;
human CXCL1 forward: 5′-AGGGAATTCACCCCAAGAAC-3′;
human CXCL1 reverse: 5′-TAACTATGGGGGATGCAGGA-3′;
human CXCL10 forward: 5′-GTGGCATTCAAGGAGTACCTC-3′;
human CXCL10 reverse: 5′-TGATGGCCTTCGATTCTGGATT-3′;
human GAPDH forward: 5′-GCACCGTCAAGGCTGAGAAC-3′;
human GAPDH reverse: 5′-TGGTGAAGACGCCAGTGGA-3′;
mouse IFNγ forward: 5′-GCCACGGCACAGTCATTGA-3′;
mouse IFNγ reverse: 5′-TGCTGATGGCCTGATTGTCTT-3′;
mouse TNFα forward: 5′-CAGGCGGTGCCTATGTCTC-3′;
mouse TNFα reverse: 5′-CGATCACCCCGAAGTTCAGTAG-3′;
mouse GAPDH forward: 5′-TGACCTCAACTACATGGTCTACA-3′;
mouse GAPDH reverse: 5′-CTTCCCATTCTCGGCCTTG-3′;
All values were normalized by GAPDH expression. The 2^−ΔΔCT method was used to analyse the mRNA expression of target genes.