BioMed Research International / 2009 / Article / Fig 2

Research Article

Bullatacin Triggered ABCB1-Overexpressing Cell Apoptosis via the Mitochondrial-Dependent Pathway

Figure 2

Apoptosis, ROS level and mitochondrial transmembrane potential were determined in KB or KBv200 cells after treatment with bullatacin. (a) Apoptotic morphology of KB and KBv200 cells was observed by Hoechst 33258 staining. The experiments were performed three times and a representative experiment is shown. (b) The apoptotic rates of KB and KBv200 cells were calculated with identified apoptosis cells by Hoechst 33258 stain dividing to accounted all cells (more than 2000 cells) after treated with bullatacin for 48 hours. Data represented means and standard errors of at least a triplicate determination. (c) Apoptotic cells were detected with Annexin V/PI after bullatacin treatment for 48 hours. The left-upper quadrant represents cells stained mainly by propidium iodide, while the right bottom quadrant represents cells stained mainly by Annexin V (apoptotic cells). The top right quadrant represents cells stained by both PI and Annexin V (secondary necrosis). The experiments were performed three times and a representative experiment is shown. (d) The apoptotic rates of KB and KBv200 cells was examined with Annexin V/PI after treated with bullatacin for 48 hours. Data represented means and standard errors of at least a triplicate determination. (e) ROS levels detection in KBv200 cells after treatment with bullatacin for 12 hours. Relative ROS levels were represented by the fold DCF intensity compared with untreated cells. The samples were duplicated and the data is a representative of three experiments. (f) Assessment of collapse in KBv200 cells. The collapse of was indicated by a reduction of intensity because the cells lost staining. Data represented means and standard errors of at least a triplicate determination.
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