Figure 5

Loss of histone deacetylase 1 and 2 (Hdacs1,2) function increases chromatin-associated histone acetylation. A. Serum-starved (SS) NIH3T3 cells were released into S-phase in the presence of DMSO or 3 μM 898. Chromatin extracts were prepared at 0 h (SS), 12 h, 18 h, and 24 h following release into S-phase for western analyses to look at changes in histone acetylation. A representative blot from three independent experiments is shown. Pan-acK, anti-pan-acetyllysine antibody. B. Quantitation for histone acetylation shown in panel A. Average intensity for a histone acetylation normalized to the total histone level were calculated from three independent experiments. C-D. Western analysis of H4K16ac using chromatin extracts prepared from NIH3T3 cells treated with DMSO or 898 (C) or 233 (D) for 24 h. E. Western analysis of H4K16ac using chromatin extracts prepared from non-targeting or Hdacs1,2-siRNA transfected cells. H4 levels serve as loading control. For panels C-E, quantitation for H4K16ac levels normalized to total H4 level from three independent experiments is shown.