Fig. 4

Cell response depends on the proteasome-ubiquitin pathway. a. JeKo1 and NCEB1 cells were treated with 1 μM etoposide, or 5 μM MG132, or both for 24 h, or vehicle, as a control, and harvested. Western blots were performed with the indicated Abs as described in the legend of Fig. 2. b. Cells were treated as in a, stained with PI and analyzed by flow cytometry. Cytometry profiles from a representative experiment are shown, the percentage of cells in the S phase is indicated on the graph. c. JeKo1 and REC1 cells were treated with 4 μg/ml etoposide (or vehicle, 0) for the indicated periods (6–24 h). Whole cell proteins were purified and analyzed by western blotting with the indicated Abs. The ratio p-eIF2α/ eIF2α estimated by densitometry, is indicated under the blots. d. Cells were treated with vehicle (−) or etoposide 4 μg/ml (+) for 24 h, then harvested. Whole cell proteins were purified, separated by SDS-PAGE, transferred onto nitrocellulose membranes and immunoblotted with an antibody against βTrcP1 protein. e. Proteins were purified from the indicated MCL cell lines and analyzed by western blot as described in d