Figure 2
Dex induces apoptosis in CEM C1-15 cells transfected with M. E4BP4: Panel A:CEM C1-15 and a mass culture CEM C1-15 cells (CEM C1-15mE*) transfected with the mouse E4BP4 expressing construct pCR3.1-mE4 were seeded at a density of 1 × 105cells/ml and treated for 72 h with either 0.1% ethanol (EtOH) or 1 μM Dex. Aliquots were taken at 24 h intervals and viable cell number was determined by trypan blue dye exclusion assay. Data represent average ± S.D. of three independent experiments with two replicates each. Panel B:A clone of CEM C1-15mE* with confirmed expression of mE4BP4 (CEM C1-15mE#3), and parental CEM C1-15 and CEM C7-14 cells were analyzed for cell viability in the presence of 0.1% ethanol or 1 μM Dex as in Panel A. Data represent averages ± S.D. of three separate experiments with two replicates each. Panel C: CEM C1-15 or CEM C1-15mE#3 cells treated for 48 h with 100 nM Dex and cell lysates were prepared from aliquots harvested every 24 h. Protein content of lysates was determined by the Bradford assay and 30 μg total protein from each sample was evaluated for PARP cleavage by Western blotting using a C-terminal-specific anti-PARP antibody (SC-7150, Santa Cruz Biotechnology).