Dmem high sugar medium

The human hepatocyte cell lines, Huh-7 and HepG2, were purchased from Cell Bank of Shanghai Institute of Life Sciences, Chinese Academy of Sciences (Shanghai China). Cells were cultured in high-sugar DMEM medium (Gibco, Gaithersburg, MD, USA) with 10% fetal bovine serum (FBS, Biological Industries, Cromwell, CT, USA) with 100 U/ml streptomycin and 100 U/ml penicillin (HyClone Company, Logan, UT, USA) in a 37°C humidified incubator with 5% CO₂.
For IL-6 treatment experiments, the Huh-7 cells were treated with 10 ng/ml IL-6 (Sigma-Aldrich, St. Louis, MO, USA) for 72 h. After the incubation, total RNA or protein was collected for miRNA array or Western blot detections.
The total RNA of Huh-7 cell was extracted, and the miRNA microarray analysis was performed by Guangzhou Ribobio Co., Ltd. (Guangdong Province, China). One microgram of total RNA was added into a nuclease-free RNA sample PCR tube, and the small RNAs (<300 nucleotides) were separated using the MilliporeSigma Centriplus Centrifugal Concentrators Microcon YM-100 (MilliporeSigma, Burlington, MA) and adding poly(A) to the 3′ end of the small RNA for hybridization at 37°C. After hybridization, dyes labeled with specific flash biotin were used, followed by scanning and analyzing the data by Affymetrix Gene Chip™ Command Console Software (AGCC).

Fetal bovine serum (Hyclone) was purchased from Thermo Scientific (Waltham, MA, USA); high-sugar DMEM medium was purchased from GIBCO (Waltham, MA, USA); penicillin and streptomycin, dimethyl sulfoxide, MTT reagent, and 0.25% trypsin digest were purchased from Solarbio company (Beijing, China); and 1× phosphate buffer (PBS) was purchased from Jiangsu biyuntian Biotechnology Research Institute (Jiangsu, China). Finally, 6 mm cell culture dishes and 6-, 12-, 24-, and 96-hole plates were purchased from Corning-Coster Company (Corning, NY, USA).