Rnaiso plus total rna kit

Total RNA was extracted from HepG2 and L02 and purified using the RNAiso Plus total RNA kit (TransGen Biotech, Dalian, China) according to the manufacturer's instruction. And then, DNase I (Takara, Dalian, China) was added to the sample to eliminate DNA contamination for RNA purity. The purity of the total RNA was assessed by measuring the absorbance ration at 260/280 nm of the samples. In addition, the quality of the RNA was confirmed by agarose gel electrophoresis. Purified RNA was reverse transcribed immediately after extraction. For qPCR experiments, HiScript® Q RT SuperMix for qPCR Kit (Vazyme, Nanjing, China) and a quantity of 1 μg total RNA were added into a 20 μl reaction volume to synthesize cDNA.

RNA was extracted from the treated and control RAW264.7 cells using RNAiso Plus total RNA kit (TransGen Biotech, Dalian, China). DNase I (Takara, Dalian, China) treatment was carried out to purify RNA. The RNA samples with OD₂₆₀/OD₂₈₀ ratios between 1.8 and 2.0 and a total amount of 0.5 μg were used for cDNA synthesis to ensure the precision of the trial. Additionally, the purification of the RNA was confirmed by agarose gel electrophoresis. The cDNA was synthesised from the RNA using reverse transcriptase and the HiScript® Q RT SuperMix for qPCR Kit (Vazyme, Nanjing, China), by following manufacturer's instructions, and stored at –20°C until use for subsequent reaction.