Journal: Cell Death & Disease
Article Title: A novel Osmium-based compound targets the mitochondria and triggers ROS-dependent apoptosis in colon carcinoma
Figure Lengend Snippet: Os affects mitochondrial function and morphology. ( a ) HCT116 cells were treated with Os (50 μ M) for 16 h, fixed and viewed under electron microscope (magnification × 25 000) White arrows pointing to mitochondria. ( b ) Relative ATP level was determined in HCT116 cells using the ATP Bioluminescence Assay Kit CLS II (Roche), as described in Materials and Methods. ( c ) A total of 6 × 10 5 HCT116 cells were incubated with 50 μ M of Os for 4 h and ETC-II activity was determined using the Human Complex activity Microplate Assay Kit (Mitosciences). ( d ) Os-treated cells were incubated with the potential-sensitive probe DiOC 6 , and ΔΨ m was analyzed by flow cytometry as described in Materials and Methods. Representative bar graphs were plotted with the G-mean values to determine the fold difference in ΔΨ m between control and Os-treated groups. ( e ) Os-treated cells were loaded with N-nonyl-acridine-orange, and cardiolipin oxidation was analyzed by flow cytometry as described in Materials and Methods. Quantification of G-mean values have been reported on the corresponding graphs. • P
Article Snippet: ATP level was determined using luciferase reagent using the ATP Bioluminescence Assay Kit CLS II (Roche, Penzberg, Germany) according to the manufacturer's instructions.
Techniques: Microscopy, ATP Bioluminescent Assay, Incubation, Activity Assay, Flow Cytometry, Cytometry