MG132 induces apoptosis in Cas-expressing cells. (A) MTS assay to determine viability of Cas-/- (EV), Cas-FL and WT MEFs treated with 1 μM MG132 and various concentrations of Z-VAD-FMK for 16 hours. (B) Apoptosis assay. Cas-FL MEFs were treated with 1 μM MG132 for different times and lysed. A total of 5 μg of protein from each lysate was used in the cell death ELISA assay (see "Methods"). Results are means ± SE from three independent experiments. (C) Cas-/- (EV), Cas-FL and WT MEFs were treated with 1 or 5 μM MG132 for 16 hours, and apoptosis was monitored as in B. (D) Cas-FL MEFs were treated for 16 hours with MG132 (upper panel) or Bortezomib (lower panel) concomitant with increasing concentrations of Z-VAD-FMK as indicated, and apoptosis was monitored as above. (E) Cell viability assay by live-dead staining. Cas-FL MEFs treated with MG132 ± 30 μM Z-VAD-FMK for 16 hours were stained with calcein (green) and ethidium homodimer (EthD, red), and images were obtained under fluorescence microscope.
Zhao and Vuori BMC Biology 2011 9:73 doi:10.1186/1741-7007-9-73