Stimulation of microglia results in histone acetylation and NF-kappaB1 p50 binding at the Nos2 promoter
. Chromatin immunoprecipitation (ChIP) was performed on microglial cell line BV2 following 3 h LPS (10 ng/ml) stimulation with or without histone acetyltransferase inhibitor C646. A Acetylated histone 4 (AcH4) ChIP on LPS-stimulated BV2. AcH4 ChIP using primers spanning 2 kb upstream of Nos2 transcriptional start site (TSS) showed that the peak of histone 4 acetylation is detected by primers amplifying −551 to −359 bases upstream of Nos2. Mean + SD. N = 3. B NF-kappaB1 p50 ChIP on LPS-stimulated BV2. p50 ChIP using primers spanning 2 kb upstream of Nos2 TSS demonstrated the peak of p50 binding at −159 to −15 bases upstream of Nos2. Mean + SD. N = 3. C The effect of pretreatment with histone acetyltransferase inhibitor C646 (C646/LPS) on NF-kappaB1 p50 binding detected by ChIP at the Nos2 promoter compared to unstimulated (VC) and vehicle treated (VC/LPS) cells. Data shown for primers amplifying −159 to −15 bases upstream of Nos2 TSS: three independent experiments (Expt).