Fig. 6: AR protein binds to cis-elements of the FSCN1 gene.

a Schema showing the AR-binding motif (ARE) obtained from JASPAR database. b Genomic location of the potential AREs on the FSCN1 gene (TSS: transcription start site). Location of the qPCR-amplified fragments (a–e) overlapping potential AREs in an 800 bp domain (little black boxes). c UCSC genome browser representations showing AR and FOXA1 binding events on FSCN1 and KLK3 loci in VCaP cells. ChIP-seq profiles indicate AR enrichment (pink and red) and FOXA1 enrichment (orange) on FSCN1 and KLK3 gene loci in the vehicle (GSM1328945 in pink), DHT-stimulated (GSM1328952 in red) or R1881-stimulated (GSM1354839 in orange) VCaP cells. d ChIP-qPCR assay confirmation of AR binding on the FSCN1 regulatory region in DHT-stimulated VCaP cells (16 h), compared with untreated cells (0 h), using either anti-AR or anti-IgG antibodies. The results are expressed in % of input for each fragment a to e. KLK3 promoter (Prom KLK3) was used as a positive control for the DHT-stimulation experiment, and an irrelevant region (Control Region) was used as a negative control. Experiments were performed with n = 3 biologically independent samples. Data were presented as mean +/− s.d. Statistical significance was determined by a two-tailed unpaired Student t test. e Schema illustrating AR signalling-mediated regulation of FSCN1 gene transcription in prostate cancer. AR Androgen receptor, DHT dihydrotestosterone, FOXA1 forkhead box A1, ADT androgen-deprivation therapy/AR antagonists, RNA PolII RNA polymerase, TF transcription factors. f ChIP-qPCR assays of FOXA1 binding on the FSCN1 regulatory region a–e in VCaP cells cultured in androgen-sensitive (left panel) or androgen-deprivation (right panel) conditions using either anti-FOXA1 or anti-IgG antibodies. The results are presented in % of input. The KLK3 promoter (Prom KLK3) was used as a positive control for the DHT-stimulation experiment. An irrelevant region (Control region) was used as a negative control. Experiments were performed with n = 3 biologically independent samples. Data were presented as mean +/− s.d. Statistical significance was determined by a two-tailed unpaired Student t test.