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. 2013 Sep 17;33(5):e00070. doi: 10.1042/BSR20130030

Figure 5. Androgen promotes interaction between AR and ARE5 in GNMT exon 1 coding region.

Figure 5

(A) A consensus ARE binding probe (lanes 1–3) and ARE5 in the human GNMT exon 1 coding region (lanes 4–12) and mutant (lanes 13,14) were incubated with nuclear extracts prepared from COS-1 cells transfected without (lanes 1, 4 and 13) or with pSG5-AR (all other lanes) in the presence of no competitor; of 50- or 100-fold molar excess of consensus, wt, or mutant cold oligonucleotide as a competitor; and rabbit polyclonal antibody anti-AR was used in the supershift assay. Arrow, AR–DNA complex; arrowhead, AR supershifted band. (B) LNCaP cells were treated with 10 nM R1881 or 0.1% ethanol for 24 h. AR binding to GNMT exon 1 ARE5 was determined by ChIP assays. Immunoprecipitated DNA (by anti-AR antibody or rabbit IgG) was purified, and regions containing ARE sites were amplified by PCR. A positive signal was obtained after R1881 treatment and ChIP with anti-AR antibodies, followed by PCR amplification using the indicated primers.