Figure 3
Rapid UPS-mediated NOXA protein turnover in MCL cells. (a) Half-life of NOXA protein in MCL cell lines (Mino, Rec1) and PHA-stimulated PBMCs of healthy donors. Cells were exposed to 20 μg/ml cycloheximide and half-life of NOXA was determined by western blot. (b) NOXA is polyubiquitinated and degraded by the proteasome. Cells were cultivated in absence or presence of 1 μM MG132 or 5 μM lactacystin for 8 h then harvest and lysed. Total polyubiquitinated proteins were isolated from cell lysates using agarose-TUBE2 beads. Protein expression and polyubiquitination state of NOXA were analyzed by western blot analysis using an NOXA-specific antibody
