Figure 4
From: Mechanisms of suberoylanilide hydroxamic acid inhibition of mammary cell growth
Effect of SAHA on histone protein and acetylation levels. (A) Equal amounts of isolated and purified histone samples from TM10 and TM2H nuclei at control (C) and treated with 2.5 μM SAHA for 24 h (T) were resolved on 15% acid-urea gel and stained with Coomassie Blue. (B) Western blot analyses of the same samples in (A) were analyzed, using 15% acid-urea gel, equilibrated to be transferred on nitrocellulose membrane, probed first with anti-H3 polyclonal antibody, striped and re-probed with anti-H4 polyclonal antibodies. Acetylated histone H4 of multiple bands (arrows) was revealed as slow migrating bands in treated versus control samples after Coomassie blue staining, while H3 showed smear of the bands attributed to an increase in both acetylation and protein synthesis. The increase in histones H3 and H4 protein and acetylation levels showed intense smeared bands after alkaline phosphatase colorimetric reaction.