Figure 7

DMC does not induce cell death in normal breast cells. (a) MCF-10A or HMECs were treated with DMC or curcumin at the indicated concentrations for 24 h. Cellular viability was assessed using calcein-AM and EthD-1. (b) MCF-10A cells or HMECs were treated with 20 μ M DMC for 16 h and observed under a phase contrast microscope. Bars, 20 μm. (c) HMECs or MDA-MB 435S cells were treated with DMC at the indicated concentrations for 24 h. Cells extracts were prepared for western blotting of the indicated proteins. Western blotting of β-actin was served as a loading control. (d) HMECs or MDA-MB 435S cells were treated with 20 μ M DMC for 16 h. Immunocytochemistry using anti-ubiquitin antibody or anti-CHOP antibody was performed and the representative images of cells are shown. Bars, 20 μm. (e) HMECs or MDA-MB 435S cells were treated with DMC at the indicated concentrations for 24 h. Cells extracts were prepared for western blotting of the indicated proteins. Western blotting of β-actin was served as a loading control. (f) HMECs or MDA-MB 435S cells were treated with or without 20 μ M DMC for 4 h and FACS analysis to detect mitochondrial superoxide using MitoSOX-Red was performed. The representative histograms are shown