C4-2 cell lines were treated with 10nM of DHT for 4 hours, followed by ChIP-Seq analyses as previously described (45 (link)). Briefly, chromatin immunoprecipitation was carried out using anti-AR antibody (Santa Cruz, Sc-816x), followed by qPCR analysis using the SYBR Green method on the QuantStudio 3 Real-time PCR system (Thermo Fisher Scientific). The primers were listed below: PSA-Enhancer (ARE3): Forward, 5′-GCCTGGATCTGAGAGAGATATCATC-3′; Reverse, 5′-ACACCTTTTTTTTTCTGGATTGTTG-3′; TMPRSS2-Enhancer (−14kb): Forward, 5′-TGGTCCTGGATGATAAAAAAAGTTT-3′; Reverse, 5′-GACATACGCCCCACAACAGA-3′.
Protocol detail
Androgen Receptor Chromatin Binding Assay
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Corresponding Organization : Harvard University
Other organizations : University of Chicago, Fred Hutch Cancer Center, University of Washington, Dana-Farber Cancer Institute
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Variable analysis
independent variables
- Treatment with 10 nM of DHT for 4 hours
- Enrichment of AR binding on the PSA-Enhancer (ARE3) and TMPRSS2-Enhancer (-14kb) regions, as measured by qPCR
- Cell line (C4-2 cells)
- Antibody used for ChIP (anti-AR antibody from Santa Cruz, Sc-816x)
- QPCR method (SYBR Green on QuantStudio 3 Real-time PCR system)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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