Figure 3
From: UGCG influences glutamine metabolism of breast cancer cells
Analysis of the redox state of MCF-7 cells. (A) The total level of reactive oxygen species (ROS) was analyzed by a fluorescence-based assay. NC = negative control (no CM-H2DCFDA), untreated = 0 μM (tert-butyl hydroperoxide = TBHP), treated = 50 μM TBHP. TBHP induces oxidative stress via several mechanisms. Data are presented as mean of n = 3 ± SEM. Unpaired t test with Welch’s correction. (B) Reduced glutathione (GSH) and oxidized glutathione disulfide (GSSG) levels were measured using a glutathione fluorometric assay kit. The ratio of GSH/GSSG was calculated. Data are presented as a mean of n = 3 ± SEM. (C) The antioxidant capacity was quantified using a colorimetric assay. Trolox was used to standardize antioxidants. Data are presented as a mean of n = 6 ± SEM. Unpaired t test. (D) Determination of reduced (NADPH) and oxidized (NADP+) form of Nicotinamide adenine dinucleotide phosphate by a luminescence-based assay. Data are presented as a mean of n = 3 ± SEM. Unpaired t test with Welch’s correction. RLU = relative luminescent units. *p ≤ 0.05, **p ≤ 0.01.
