Anti di methyl histone h3 lysine 4

ChIP analysis of transcriptionally active chromatin markers interacting with hTERT promoter was performed using the EZ-ChIP kit (Upstate Biotechnology) according to the instructions included in the kit. ChIP-validated antibodies used were: anti-acetyl-histone H3 lysine 9, anti-acetyl-histone H4, anti-tri-methyl histone H3 lysine 9 and anti-di-methyl histone H3 lysine 4, all from Millipore. ChIP-purified DNA from control cells (untreated) and cells treated with CDDO-Me (0.125–0.5 μM) for 5 days was amplified by PCR using hTERT promoter primers: forward, 5′-TCCCCTTCACGTCCGGCATT-3′; reverse, 5′-AGCGGAGAGAGGTCGAATCG-3′. The PCR products were separated on 2% agarose gel electrophoresis and visualized by ethidium bromide staining. The hTERT primers amplified a DNA fragment of 200 bp.