Figure 6. MOR-induced binding of NF-κB to the enhancer region of the CCL2 promoter in vivo.

Chromatin was prepared from human PBMCs and digested into one to two nucleosome fragments that were used for ChIP analysis. Chromatin (15–30 μg) was incubated with protein A magnetic beads and an anti-acetyl-H3 (Anti-ac-H3) antibody or an anti-p65 antibody overnight. Protein/DNA complexes were then washed and separated. Purified DNA was then subjected to PCR analysis for GAPDH or the CCL2 enhancer region (ER).