Figure 6

HDAC inhibition induced enhanced binding of acetylated histone H3, CBP/p300 and CREB to NKG2D-ligand promoters. (a) Phospho-kinase profiler array of HEK-293 cells pretreated with or without 10 μM C646 for 3 h and incubated with 100 nM LBH589 for one additional hour, followed by lysis of the cells. Detection of phosphorylated kinases was performed with a digital ECL imager. (b) Chromatin immunoprecipitation (ChIP) of HEK-293 cells treated with 100 nM LBH589 for 3 h. Pull down was performed with antibodies against acetylated histone H3, CBP (also binding to p300) and CREB. Precipitated promoter sequences were detected by real-time PCR and calculation was implemented using the % input method. Values were normalized to RPL30.