Cistrome analyses of AR-V7.

Although AR-V7 has been intensively studied, it remains unclear whether AR-V7 can specifically activate a distinctive transcriptional program from the full-length AR (AR-FL), and whether AR-V7 may play a role in accelerating the metastatic progression of castration-resistant PCa (CRPC). In this study, we hypothesize that AR-V7 can drive a distinct transcription program from AR-FL in CRPC condition. To test this, we used LNCaP model with inducible overexpression of AR-V7 or AR-FL to examine the effects of AR-V7 overexpression or AR-FL overexpression stimulated with DHT for 4 hours on AR-V7 or AR-FL cistromes. We also studied the effects of AR-V7 on cistromes of pioneer factor FOXA1 and active histone marker H3K27ac, and the function of phosphorylation at Ser81 on AR-V7 function in CRPC. Overall design: ChIP-seq analyses of V5 tagged AR-V7, AR-C, FOXA1 and H3K27ac were performed to examine AR-V7, AR-C, FOXA1 and H3K27ac binding with or without overexpression of AR-V7 or AR-V7-S81A in LNCaP stable cell line under 5% charcoal-stripped FBS condition. The stable cells were generated by stably infecting LNCaP cells with tetracycline inducible overexpression of wild-type AR-V7 or S81A-AR-V7 virus. These stable cells were grown in the medium containing 10% tetracycline-free FBS.