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Figure 2 | Molecular Cancer

Figure 2

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

Figure 2

Autophagic cell-death by rottlerin in breast CSCs. (a) Redistribution of pEGFP-LC3. Breast CSCs were stably transfected with a pEGFP-LC3 fusion construct and cultured in complete stem cell culture medium, and treated with Rott (0, 0.5, 1 and 2 μM) for 48 h. Cells were visualized under a fluorescence microscope to examine the expression of LC3-II. LC3 expression increases by increasing Rott concentration in breast CSCs. (b) Electron microscopy shows the ultrastructure of breast CSCs treated with different concentrations of Rott (0 and 2 μM) in complete stem cell culture medium for 48 h. Arrows indicate autophagosomes including residual digested material (red arrow). (c) Punctate dot quantification in pEGFP-LC3-positive breast CSCs treated with Rott (0, 0.5, 1 and 2 μM) for 48 h. Quantification of puncatate dot per cell based on number of punctuate dot in pEGFP-LC3-positive cells. Quantification represents at least 100 cells counted and scored per treatment. (d, e and f) Punctate dot quantification in pEGFP-LC3-positive breast CSCs co-treated with Rott (2 μM) and Baf (10 nM), 3-MA (10 mM) or CHX (10 μg/ml) for 48 h. Quantification of puncatate dot per cell based on number of punctuate dot in pEGFP-LC3-positive cells. Quantification represents at least 100 cells counted and scored per treatment. 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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