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Figure 7 | Molecular Cancer

Figure 7

From: Rottlerin-induced autophagy leads to the apoptosis in breast cancer stem cells: molecular mechanisms

Figure 7

Apoptotic cell-death measured by annexin-V/PI staining in breast CSCs. Cells were co-treated with Rott (2 μM) and Baf (10 nM), 3-MA (10 mM) or CHX (10 μg/ml) (a) Rott (2 μM) and Baf (10 nM), (b) Rott (2 μM) and CHX (10 μg/ml), and (c) Rott (2 μM) and 3-MA (10 mM) in complete stem cell culture medium for 48 h. Apoptosis was measured by annexin-V/PI staining followed by flow cytometry. Data are the means of triplicate experiments. Representative histograms are shown of co-treated with (a) Rott (2 μM) and Baf (10 nM), (b) Rott (2 μM) and CHX (10 μg/ml), and (c) Rott (2 μM) and 3-MA (10 mM) breast CSCs stained with annexin-V and propidium iodide. After 48 h of culture, three populations of cells were observed: viable cells (negatively stained, lower left quadrants); early apoptotic cells (annexin-V positive and propidium iodide negative, lower right quadrant) and cells in the late stages of apoptosis (annexin-V and propidium iodide positive, upper right quadrants). By increasing Rott concentration at 48 h, a greater number of breast CSCs underwent the early and late stages of apoptosis. (d) Kinetic evaluation of cells undergoing the early stage of apoptosis in breast CSCs. Cells staining with annexin-V only, showing the early stage of apoptosis (% early apoptosis) (e) Kinetic evaluation of cells undergoing the early stage of apoptosis in breast CSCs. Cells staining with annexin-V alone and with both annexin-V and propidium iodide were combined, giving the total number of cells at both the early and late stages of apoptosis (% cell death). Data are reported as the mean ± standard error (SE) of percentage of cells. n = 5, *P < 0.05 when compared with Rott treated in an identical manner.

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