Gene Expression Analysis in HepG2 Cells

Total RNA was extracted from HepG2 cells using RNAios Plus. The concentration of RNA was determined by measuring the absorbance at 260 nm. Subsequently, cDNA was synthesized using the PrimeScript™ RT reagent kit. Samples were prepared using TB Green® Pre-mix Ex Taq™ II, and quantitative real-time PCR was conducted on a real-time Thermal cycler 5100 (Thermo Fisher Scientific, CA, USA). The relative concentrations of mRNA were normalized to the expression levels of GAPDH to quantify gene expression. Data analysis was performed using the 2^−△△CT method [36 (link)]. The primer sequences utilized in the PCR are provided in Supplementary Table S1.