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. Author manuscript; available in PMC: 2014 Mar 9.
Published in final edited form as: Mol Cell. 2007 Aug 3;27(3):380–392. doi: 10.1016/j.molcel.2007.05.041

Figure 3. Characterization of TMPRSS2-Regulatory Regions.

Figure 3

(A) ChIP analyses of AR occupancy on the PSA and B39 in VCaP cells. VCaP cells (a prostate cancer cell line that expresses TMPRSS2:ERG fusion gene) were treated with 100 nM DHT for 1 hr. ChIP assays were performed with an anti-AR antibody.

(B) Spatial communication between the TMPRSS2 enhancer and promoter. 5C was performed using fixed or EcoRI- or BtgI-digested chromatin from vehicle- or DHT-treated LNCaP cells. Primers (Table S1) flanking the –13.5 kb or 462 kb AR binding region and –700 bp promoter region were used to PCR amplify DNA after ligation. Control PCR was performed using chromatin before restriction enzyme digestion.

(C) Schematic representation of the TMPRSS2 14 kb upstream regulatory region. Potential five ARE clusters and fourteen 1 kb fragments are shown.

(D) ChIP analyses of AR recruitment to five potential ARE regions. ChIP assays were performed with anti-AR antibodies in LNCaP cells treated with vehicle or 100 nM DHT for 4 or 16 hr.

(E) Systematic mapping of enhancer elements within the TMPRSS2 14 kb upstream regulatory region. Fourteen 1 kb TMPRSS2 upstream sequences were subcloned into the pGL3-promoter vector and transfected into LNCaP cells. Cells were treated with vehicle or 100 nM DHT for 24 hr. The data are presented as the mean ± SE of two to three replicates, (A), (D), and (E).