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. 2017 Feb 21;7:42957. doi: 10.1038/srep42957

Figure 6. PST Analogs Act on Cancer Cell Mitochondria and Cause Mitochondrial Dysfunction.

Figure 6

(a) H2DCFDA was used to measure whole cell ROS in MV-4-11 and U-937 cells treated for 3 hours with image-based cytometry. *p < 0.01 vs. DMSO control. (b) The MitoXpress® Xtra - Oxygen Consumption Assay was used to monitor oxygen consumption via fluorescence generation. Cells were treated, and the fluorescent MitoXpress® reagent was added and monitored at Ex. 380 nm and Em. 650, every 2 minutes for 2 hours at 37 °C. Oxygen consumption rates were calculated by measuring the slopes of the linear regions of the oxygen consumption curves. *p < 0.001 vs. DMSO control. (c) Western blot analysis of E6-1 following treatment with the indicated drugs for 6 hours. Results are representative of 3 independent trials. (d) Detection of ATP levels following treatment with SVTH-5, -6, and -7, PST, and Taxol using the luciferase-luciferin ATP determination assay. Amount of ATP was expressed as number of moles of ATP over micrograms of protein. Results are shown as the mean ± SD from at least 3 independent experiments. *p < 0.05 vs DMSO control. (e) Western Blot analysis of Cyto c release (of post mitochondrial supernatant) from directly treated mitochondria isolated from MV-4-11 cells for 2 hours. SDHA was probed in the mitochondrial pellet samples as loading controls. All quantitative values are expressed as mean ± SD from at least 3 independent experiments. Western blots are representative of at least 3 independent experiments.