High glucose dulbecco s modified eagle media

The human HCC cell lines PLC, Hep3B, SMMC-7721, Huh7, HepG2, the normal liver cell line L02 and embryonic kidney cell line HEK-293T were purchased from the Cell Bank of Typical Culture Preservation Committee of Chinese Academy of Science (Shanghai, China). Human HCC cell lines MHCC-97H and HCC-LM3 were established at the Liver Cancer Institute, Zhongshan Hospital, Fudan University. These cells were stored in liquid nitrogen and cultured in 5% CO2 at 37°C with high glucose Dulbecco's modified Eagle media (GIBCO, Grand Island, NY) supplemented with 10% FBS (GIBCO).
The negative control small interference RNA (siNC, 5′-UUCUCCGAACGUGUCACGUTT-3′), and Nrf2 siRNA (5′-CAUUGAUGUUUCUGAUCUATT-3′) were synthesized and inserted into the LV3-GFP vector (named as LV3-shCon or LV3-shNrf2). For Nrf2 overexpression, the Nrf2 expression plasmid was cloned into LV5-GFP vector (named as LV5-Nrf2). Thereafter, HEK-293T cells (1 × 10⁶ cells) were transfected with the respective plasmid (LV3-shCon, or LV3-shNrf2, or LV5-NC, or LV5-Nrf2) and packaging vectors by GenePharma Co. Ltd (Shanghai,China). HCC cells were then infected with the packaged lentivirus under medium culture.

Human HCC cell lines SK-HEP-1 and Huh7 were purchased from the Cell Bank of Chinese Academy of Sciences, Kunming. HCC cell line MHCC97-H was purchased from FuDan IBS Cell Center (FDCC), Shanghai. Cells were cultured in 5% CO2 at 37 °C with high glucose Dulbecco’s modified Eagle media (Gibco, USA) supplemented with penicillin (100 IU/mL), streptomycin (100 mg/mL) and 10% FBS (Gibco, USA).