(A) Time course ChIP assay for cluster 2 genes. LNCaP cells (150mm dishes) were transfected with either siControl (LowGC) or siHOXB13 (siHOX71) for 48 h prior to the addition of R1881 (100nM). Following hormone addition (1,2,4, and 8 h), cells were fixed with formaldehyde and sheared chromatin was immunoprecipitated with the indicated antibodies (IgG, AR, TRAP220, P300, SRC1). Real-time PCR was used to assess the amount of DNA immunoprecipitated in response to hormone treatment, presented as a percentage of input chromatin. The position of the AR binding sites (IR3 or IR8) identified by bioinformatic analysis in relation to the transcription start site (TSS) is provided in the gene structure illustrations. Error presented as +/− SD of triplicate points. Veh; vehicle.
(B) Time course ChIP assay for cluster 3 genes. Experiments were performed same as in (A). Data for (A) and (B) are colored according to gene cluster as presented in Figure 2.