Fig. 14.1.

Dose-dependent and differential effects of hydroxamate H6CAHA and mercaptoacetamide 6MAQH on histone H4 and α-tubulin acetylation levels in MCF7 cells were analyzed after 4-h treatment with indicated concentrations of HDAC inhibitors. Cells lysates were prepared in a lysis buffer containing 20-mM Tris–HCl (pH 7.4), 2-mM EDTA (pH 7.4), 2-mM EGTA (pH 7.4), 6-mM mercaptoethanol, 10 μg/ml leupeptin, 2 μg/ml aprotinin, and 1% NP40. The samples were subjected to SDS-polyacrylamide gel electrophoresis. The indicated proteins were probed with a polyclonal rabbit antibody against acetylated histone H4 or α-tubulin (Upstate, Inc). The immune-reactive bands were detected by enhanced chemiluminescence (Amersham Biosciences)