Research Paper Volume 16, Issue 11 pp 9334—9349

Figure 4. BNIP3(L)-high MCF7 cells show higher ATP levels and higher anti-oxidant capacity; however, mitochondrial mass remains unchanged. MCF7 cells stably-transduced with the BNIP3(L)-GFP constructs were subjected to FACS sorting, to isolate the 5% highest GFP (GFP-high) and the 5% lowest GFP (GFP-low) sub-populations. (A) The GFP-high and GFP-low subpopulations were plated in complete DMEM medium and incubated with the Cell-Titer-Glo 2.0 Reagent for 15 min to determinate the ATP levels. Data are shown as the mean ± standard deviation (SD) (n = 3). Statistical significance was determined using an unpaired Student’s t-test, * p ≤ 0.05, ** p ≤ 0.01. (B) Mitochondrial mass was assessed using MitoTracker Deep Red by flow cytometry. Data are shown as mean ± standard deviation (SD) (n = 3). Statistical significance was determined using an unpaired Student’s t-test, n.s. not statistically significant. (C) The anti-oxidant capacity was measured with the GSH/GSSG-Glo Assay. Data are shown as the mean ± standard deviation (SD) (n = 3). Statistical significance was determined using an unpaired Student’s t-test, ** p ≤ 0.01, ***p ≤ 0.001.