4.4. Exogenous Induction of Nrf2 by Quercetin

Stock solutions of Que were freshly prepared by dissolving it in Dimethyl sulfoxide (DMSO). The stock solutions were subsequently diluted with Dulbecco’s Modified Eagle Medium: Nutrient Mixture F-12 (DMEM/F12-HAM) media. The final concentrations of DMSO in the medium were ≤0.01% (v/v). Appropriate controls with only the vehicle DMSO were included in all experiments. For exogenous Nrf2 modulation, the appropriate and non-toxic dose of Que was determined. For that, sub-confluent granulosa cells were treated with different concentrations of Que (0, 10, 20, 50, 100 or 200 µM) for 24 h. Treated cells were then subjected to intracellular ROS level detection, cell cycle assay, cell proliferation assay and assessment of mitochondrial activity. Moreover, mRNA and protein expression patterns of candidate genes were quantified in each treatment group.