Figure 6.

TNFα-Responsive Enrichment of NF-κB, B-myb, HDAC1, SMRT, and mSin3A at the Chromatin Region Containing Site I and Site II of the AR Promoter in LNCaP Cells

Soluble chromatin fragments (average, ∼500 bp) from LNCaP cells treated with TNFα (for 30 min or 60 min), or with vehicle (PBS), were analyzed by ChIP. A, The immunoprecipitated DNAs were amplified with AR-specific PCR primers (forward at −38; reverse at +246). These primers are described in Materials and Methods. B, PCR of the immunoprecipitated DNAs using upstream primers corresponding to the AR promoter from −760 to −460 (negative control). C, ChIP using antibodies to acetylated histones and to phospho-RNA polymerase II. AR-specific primers as in panel A were used in PCR amplification. Anti-Cox2 served as a negative control antibody. Amplified PCR products of 1% input DNAs from cells with or without TNFα treatment are shown. Ac-histone, Acetylated histone; Pol II, polymerase II.