Figure 1

Rottlerin inhibited cell viability, and induced apoptosis and cytoplasmic vacuolation in breast CSCs. (a) Cells were grown in complete stem cell culture medium and treated with Rott (0, 0.5, 1 and 2 μM) for different time points. Cell viability was measured by XTT assay. Blue color (12 h), red (24 h), green (48 h), and violet (72 h). (b) Breast CSCs were treated with Rott (0, 0.5, 1 and 2 μM) for 48 h, and the autophagic vacuoles were counted under phase contrast microscope. (c) Breast CSCs were co-treated with Rott (2 μM) and Baf (10 nM), 3-MA (10 mM) or CHX (10 μg/ml) for 48 h, and the autophagic vacuoles were counted under phase contrast microscope. (d) Breast CSCs were cultured in complete stem cell culture medium and treated with the different concentrations of Rott (0, 0.5, 1 and 2 μM) for 48 h. Representative images were obtained by phase contrast microscopy. Magnification, 20X. Red arrows indicate cytoplasmic vacuoles developed by the effect of different concentration of Rott (0, 0.5, 1 and 2 μM). (e) Breast CSCs were cultured in complete stem cell culture medium and co-treated with Rott (2 μM) and Baf (10 nM), 3-MA (10 mM) or CHX (10 μg/ml) for 48 h. Representative images were obtained by phase contrast microscopy. Magnification, 20X. Red arrows indicate cytoplasmic vacuoles. 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.