ChIP assays of binding of NFκB p65, ERβ and AcH3 to the MCP-1 and CINC-2β promoters.

Cells were pretreated without or with E2 for 24 hrs, then stimulated with TNF-α (1 ng/mL) for 1 hr. ChIP samples were prepared as described in the text and analyzed using antibodies specific for p65, ERβ or AcH3. The immunoprecipitated DNA fragments and input DNA were analyzed by by real-time PCR. The y axis shows values were normalized to input DNA with values for vehicle treatment defined as 1. The numbers represent result from three experiments repeated in duplicate. *p<0.05 vs. Vehicle-treated RASMCs; #p<0.05 vs. TNF-α-treated RASMCs.