Dmem f12 1 1 medium

Two human cancer cell lines, MCF7 (ER⁺ breast cancer adenocarcinoma) and Hela (cervix adenocarcinoma) were used for transient transfection experiments. MCF7 cells were grown in DMEM-F12 (1:1) medium (Wisent, St-Bruno, QC, Canada) supplemented with 5% fetal bovine serum (FBS), penicillin/streptomycin (100 IU/mL-1%, w/v), Hepes and 1 nM estradiol. HeLa cells were grown in Eagle’s Minimum Essential Medium (Wisent, St-Bruno, QC, Canada) enriched with 5% FBS, 1% L-glutamine and penicillin/streptomycin (100 IU/mL−C1%, w/v). Both cell lines were grown at 37 °CC in a 5% CO₂ atmosphere and 95% relative humidity.

The ALM12 cell (mouse normal hepatocyte) line was purchased from the BioVector NTCC Typical Culture Preservation Center (SCSP-550, Beijing, China). The cells were cultured in a DMEM/F-12 (1:1) medium (WISENT) containing ITS Liquid Media Supplement (WISENT, 315-081-XL), 10% (v/v) fetal bovine serum (Gibco, Grand Island, NY, USA), 100 U/mL penicillin, and 100 μg/mL streptomycin (Sigma, Shanghai, China) for three days at 37 °C with 5% CO₂. AML12 cells were pretreated with DHA (50 μM, 53171, Sigma, Shanghai, China) for 12 h, followed with H₂O₂ (400 μM) challenging for 2 h. The cells collected were used in a cell viability assay, along with immunofluorescence, Western blotting, and flow cytometry tests.