Figure 4

Dex-mediated upregulation of Bim and E4BP4 is restored in CEM C1-15mE#3 cells: Panels A and C: CEM C7-14, CEM C1-15 and CEM C1-15mE#3 cells were treated with 0.1% ethanol or 1 μM Dex for 24 h and total RNA was extracted in TriZol. Seven microgram RNA was subjected to reverse transcription reaction and real-time qPCR using primers specific for Bim or Puma (Panel A) or E4BP4 (Panel C) as listed in Table 1. Fold change in expression of each transcript by 1 μM Dex was calculated by the Pfaffl method using β-actin as a reference. Data represent averages ± S.D. from three independent experiments, which were analyzed for statistical significance using a paired student t-test, as shown in the table below each bar chart. Shaded boxes represent significant differences with p-values ≤ 0.05. Panels B and D: CEM C7-14, CEM C1-15 and CEM C1-15mE#3 cells were treated with 0.1% ethanol (E) or 100 nM Dex (D) for 24 h and total protein was extracted in lysis buffer. Protein content of lysates was measured by the Bradford assay, and 30 μg protein from each sample was evaluated by Western blotting for Bim and Puma expression (Panel B) or for E4BP4 expression using two different antibodies from Santa Cruz Biotechnology (Panel D). An antibody for GAPDH was used as a reference.