Salinomycin Induces Autophagy in Colon and Breast Cancer Cells with Concomitant Generation of Reactive Oxygen Species
Figure 3
Impact of salinomycin on caspase activation.
Caspase activities were measured after treatment of breast and colon cancer cell lines with salinomycin for 28 hours. (A) Caspase 9 activity measured with Z-LEHD-aminoluciferin as a substrate; (B) Caspase 8 activity measured with Z-LETD-aminoluciferin as a substrate; (C) Caspase 3/7 activity measured in colon cancer cell lines with Z-DEVD-aminoluciferin as a substrate. Measurements were in MCF-7 cells were omitted as these cells are caspase 3-negative. Significant increases in measured luminescence in comparison to the untreated cells are indicated in the graphs (*: p<0.05; **: p<0.01, ***: p<0.001 by t-test). RKO cells were most sensitive to salinomycin treatment for all the measured caspases. (D) Western blot analysis of caspase status. Procaspase 3 and procaspase 9 expression were determined in SW480, SW620, RKO, and MCF-7 cells. β-Actin was used as a loading control. (E) Caspase 3 status in T47D and MDA-MB-453 cells. Procaspase 3 expression was determined by Western blot; α-Tubulin was used as a loading control. (F) Caspase 3/7 activity measured in breast cancer cell lines with Z-DEVD-aminoluciferin as a substrate. Caspase 3 activity was not measured in MCF-7 as these cells are caspase 3-negative.