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AR NeuroD HNF4alpha1 OCT Octamer Vsx1 Irx Nkx2 HOXA7 ISL1 Dobox4 Lhx9 IPF1 ERR1 TATA CREB Arx PU1 E2F STAT Poly GLI1 SOX10 Msx Lbx2 YY1 E47 core GKLF ATF1 ZBRK1 Evi ER Obox6 IRF SMAD Tax TBX5 Barx CDP SZF1 AML1 HEB SREBP POU3F2 ALX ARP Xvent GTF2IRD1 RFX HNF1 Bach2 ZNF515 Lmo2 LF Cdx HSF2 IRF8 cebpe AIRE Meis1 MRG2 c BCL6 MZF1 XFD GATA DMRT3 FOX HMGIY RSRFC4 HLF NF RelB AhR LEF1 BRCA1 MEF Dbx AP Sp1 PLZF APOLYA Vax AREB6 Elf LXR
C AR mRNA Relative expression (2 ǻǻCT )
chr1
Random ARBS Most Lost Most maintained
B
Enhanced
Bicalutamide
A
1
shNT shAR
* 0.5 LNCaP-Hes6HA shNT
shAR
ĮAR ĮTubulin
Figure S4, related to Figure 2. Hes6 overexpression induces variable changes in AR chromatin binding. (A) Representative examples are given of ChIPseq tracks showing an ARBS at MTOR that is enhanced by Hes6 in bicalutamide and two ARBS at FKBP5 that are lost despite Hes6 in bicalutamide (indicated with an arrow) and one that is rescued (indicated with a star). The rescued ARBS was used to design the primers used for Fig 2F. (B) Heatmap showing relative enrichment of transcription factors by MEME (http://meme.nbcr.net/meme/) when comparing the 1000 mostmaintained ARBS, the 1000 mostlost and a random matched-size set across all ARBS. (C) Hes6-overexpressing LNCaP cells were transduced with a pSicoR lentivirus to stably knockdown AR (40% knockdown). AR mRNA levels measured by q-PCR and one representative western blot are shown; n = 3, error bars represent mean ± SEM; *p = 0.0002 by t-test.
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