Figure 3. Vorinostat alone or in combination with mithramycin A increases global as well as MICA and MICB promoter-specific histone H3 Lysine 9 (H3K9) acetylation in MCC cell lines.
(a) Chromatin immunoprecipitation (ChIP) assay was performed with untreated, vorinostat (V), mithramycin A (MA), or the combination thereof (V+MA) treated WaGa cells followed by a qRT-PCR using MICA or MICB promoter specific primers. CT values of anti-acetyl-H3K9 (AcH3K9) antibody or rabbit IgG isotype control precipitated DNA were normalized to total histone H3 antibody as described in materials and methods. White bars represent the percentage of acetylated H3K9, grey bars the respective control. Experiments were performed in duplicates and results are expressed as mean ± SEM. (b) Global H3K9 acetylation of untreated, V, MA, or V+MA treated MCC cell lines was determined by immunoblot with the same AcH3K9 antibody used in the ChIP assay; β-tubulin served as loading control.