Carnosol Induces ROS-Mediated Beclin1-Independent Autophagy and Apoptosis in Triple Negative Breast Cancer
Figure 3
Induction of caspases-mediated apoptosis by carnosol in the MDA-MB-231 cells.
(A) Carnosol induced apoptosis in the MDA-MB-231 cells. Annexin V binding was carried out using Annexin V & Dead Cell kit (Millipore). Cells were treated with DMSO or various concentration of carnosol for 24 h. Detached and adherent cells were collected and stained with Annexin V and 7-AAD and then the events for early and late apoptotic cells were counted with the Muse Cell Analyzer as described in Materials and Methods. (B) Western blot analysis of caspase 3, -9, and -8 activation, PARP cleavage and Bcl1 and Bax expression in MDA-MB-231 cells were treated with increasing concentrations of carnosol (25, 50 and 100 µM) for 24 h. (C) Carnosol induces the depolarization of mitochondrial membrane. Mitochondrial membrane potential (MMP) was assessed with the Muse Cell Analyzer using the Muse MitoPotential kit as described in Materials and Methods. Data represent the mean ±SEM of at least 3 independent experiments. Student's t test was performed to determine the significance (*p<0.05, **p<0.01 and ***p<0.005).