Figures & data
Figure 1. Panobinostat posttranscriptionally enhances Dicer protein expression. A. JAR cells treated with increasing doses of Panobinostat for 24 h were harvested, lysed, and probed for expression of Dicer, Argonaute-2, acetylated histone 3, Caspase-3, cleaved Caspase-3, and β-actin via Western blot. Data represent three independent experiments. B. Data from A quantified from three independent experiments. C. RNA was harvested from the previous cell treatments and Dicer and β-actin mRNA levels were assessed via quantitative real-time RT-PCR. D. JAR cells treated with 25 nM Trichostatin A were harvested and probed for expression of Dicer, acetylated histone 3, and β-actin via Western blot. The data represent three independent experiments. E. Data from D quantified from three independent experiments. Error bars ± SEM, *P < 0.05, **P < 0.005, ***P < 0.0005.
Figure 2. Dicer protein expression over 48 h of Panobinostat treatment. A. JAR cells treated with 25 nM Panobinostat over the course of 48 h were harvested and probed for expression of Dicer, acetylated histone 3, and β-actin via Western blot. B. JAR cells treated with 25 nM Trichostatin A over the course of 48 h were harvested and probed for expression of Dicer, acetylated histone 3, and GAPDH via Western blot. The data represent two independent experiments.
Figure 3. Panobinostat reduces the proteasomal degradation of Dicer. A. JAR cells treated for 24 h with Bortezomib, Panobinostat, or Bortezomib + Panobinostat were harvested and probed for expression of Dicer, acetylated histone 3, Caspase-3, cleaved Caspase-3, and β-actin via Western blot. The data represent three independent experiments. B. Data from A quantified from three independent experiments combined and the statistics are displayed in the table. Error bars ± SEM, *P < 0.05, **P < 0.005.
Figure 4. Panobinostat decreases Dicer activity. A. Protein lysates (20 µg) from JAR cells +/− 24 h of Panobinostat treatment were used in the Dicer activity assay and the amount of pre-miR-124a and mature-miR-124a were quantified. B. Dicer's relative specific activity was calculated using the ratio of mature-miR-124a produced to Dicer protein expression in a lysate. The data represent three independent experiments. Error bars ± SEM, **P < 0.005.
Figure 5. Bortezomib decreases Dicer activity. A. 20 µg of protein lysates from JAR cells +/− 24 h of Bortezomib treatment were used in the Dicer activity assay and the amount of pre-miR-124a and mature-miR-124a were quantified. B. Dicer's relative specific activity was calculated using the ratio of mature-miR-124a produced to Dicer protein expression in a lysate. The data represent three independent experiments. Error bars ± SEM, **P < 0.005.
