Cas is required for proteasome inhibition-induced cell death. (A) Cas-/- (EV) and Cas-FL cells cultured in 60 mm dishes were treated with vehicle, or MG132, or Bortezomib overnight, and the images were obtained under phase contrast microscope. Scale bar, 25 μm. (B) Cell viability measurement by MTS assay. MEFs grown in 96-well plates were treated with different concentrations of the proteasome inhibitors MG132 or Bortezomib for 16 hours. A total of 20 μl of MTS reagent was added to each well and incubated for another two hours. Absorbance at 490 nm was detected on a plate reader. The data presented depict the MEAN ± SE from three independent experiments. (C and D), 293T cells (C) or HeLa (D) cells transiently transfected with Cas shRNAs, control shRNA or empty vector, were treated with MG132 or Bortezomib for 16 hours, then assayed and results are presented as in panel A. shRNA-mediated reduction of Cas expression is shown in immunoblot.
Zhao and Vuori BMC Biology 2011 9:73 doi:10.1186/1741-7007-9-73