Figure 6

Resistant ovarian cancer C200 cells have the ability to switch between energy states. (A) Equal number of cells were plated in multi-well Seahorse Biosciences XF96 plate (Boston, MA), and after calibration time indicated doses of 2-deoxyglucose (2DG) were injected and measurements of OCR and ECAR were recorded in triplicates. (B) Bar graph representation of the dose-dependent effect on ECAR and OCR after 2DG inhibition of glycolysis. ***p < 0.001, **p < 0.01, *p < 0.05 2DG treated compared to untreated. (C) Equal number of cells were plated in multi-well Seahorse Biosciences XF96 plate, and after calibration time indicated doses of oligomycin were injected and measurements of OCR and ECAR were recorded in triplicates. (D) Bar graph representation of the dose-dependent effect on ECAR and OCR after oligomycin inhibition of mitochondrial function. ***p < 0.001, **p < 0.01, *p < 0.05 oligomycin treated compared to untreated. All Seahorse experiments were performed in triplicates and replicated twice. A2780 and C200 were plated at 4000 cells per well in 96 wells and treated with indicated concentrations of (E) 2DG or (F) oligomycin and live cells were measured at 48 h using Promega Cell Viability Assay. The data is represented as percent survival compared to untreated control cells, which were taken as 100% live cells. **p < 0.01, *p < 0.05 C200 compared to A2780. The experiment was performed in triplicates thrice. Chemoresistant C200 (G) and PEO4 (H) cells were treated with cisplatin in the presence or absence of 2DG or oligomycin and live cells were measured at 48 h using Promega Cell Viability Assay. The data is represented as percent survival compared to untreated control cells, which were taken as 100% live cells. The cell survival studies were carried out in triplicates and replicated thrice. ***p < 0.001, **p < 0.01, *p < 0.05 combination compared to individual treatments.