Fig. 5: Mitochondrial effects of BMS.
From: Synergistic targeting of cancer cells through simultaneous inhibition of key metabolic enzymes
A The indicated EV control and KRASG12V/MYC cancer cells were transduced to stably express COX8-EGFP, enabling visualization of mitochondria. Cells were treated for 2 days with indicated conditions, followed by fixation and confocal imaging. High-resolution 3D rendered mitochondria of representative cells are displayed. Scale bar = 10 µm xy-plane. B The number of mitochondria per cell and C mitochondrial mean branch length was quantified, n = 3, one-way ANOVA with Šídák’s multiple comparisons test. D KRASG12V/MYC cells were treated as indicated, followed by analysis of mitochondrial structure via transmission electron microscopy. Scale bar = 500 nm. The rectangular selections are shown in the lower row at a higher magnification. E KRASG12V/MYC cancer cells were treated with GNE and/or BMS for 4 h, followed by Seahorse metabolic flux analysis. Oligomycin, FCCP and Rotenone/Antimycin A were injected into the wells at the indicated timepoints, followed by quantification of the oxygen consumption rate (OCR). F The experiment was performed as in E for quantification of the extracellular acidification rate (ECAR). Seahorse experiments show mean ± SD, n = 4. G Energy map summarizing the Seahorse experiments and annotating the different treatments to metabolic states (Aerobic, Glycolytic, Energetic or Quiescent). H The indicated cell lines were treated for 4 days with GNE and/or BMS and the extracellular pH was determined. Statistical analysis was done using two-way ANOVA with Dunnett’s multiple comparisons test.
