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. Author manuscript; available in PMC: 2016 Jun 8.
Published in final edited form as: Cancer Cell. 2015 Jun 8;27(6):797–808. doi: 10.1016/j.ccell.2015.05.005

Figure 5. ERG maintains YAP1 expression in human prostate cancer cells.

Figure 5

(A-C) Significant concordance between ERG and YAP1/TAZ gene expression programs in VCaP cells. Gene expression determined by RNA-Seq of VCaP cells transiently transfected with non-targeting siRNA (siCtrl, n=2), siRNAs targeting ERG (siERG, n=2, using 2 independent siRNA oligos) and siRNAs targeting YAP1/TAZ (siYAP1/TAZ, n=2, using 2 independent siRNA oligo mixtures). (A) Heat-map; (B) overlap between significantly (Q<0.05) upregulated and downregulated genes; and (C) GSEA analysis of overlap between siERG-mediated gene expression changes and siYAP/TAZ downregulated genes. Two tailed Chi-square with Yates correction in B.

(D) Western blot (WB) analysis of VCaP cells transfected with indicated siRNA oligos. Numbers indicate relative expression values, with value in siCtrl adjusted to 1.

(E) Highly significant overlap between positions of TEAD4 and ERG peaks in VCaP cells (hypergeometric p-value=0 calculated by Bioconductor package “ChIPPeakAnno”).

(F) Tendency toward close juxtaposition of ERG (A/C)GGAA(G/A) and TEAD4 GGAAT(G/T)(T/C) recognition motifs in ERG-TEAD4 overlapping peaks in VCaP cells.

(G) UCSC genome browser views of ChIP-Seq data from VCaP cells using IgG, anti-ERG and anti-TEAD4 antibodies.

(H) qPCR analysis of ChIP experiments from siCtrl, siERG-1 or siERG-2 oligo transfected VCaP cells using indicated antibodies. Data represent mean±standard deviation (n = 4) from one of three independent experiments. Student's t test.

(I) Significant correlation between ERG expression and presence of nuclear YAP1 in primary human prostate tumors. Immunohistochemical (IHC) staining of prostate cancers (PC1 and PC2) with ERG and YAP1 antibodies. Note, YAP1 is strongly expressed in benign basal but not benign luminal epithelial cells (arrows), but is present in neoplastic luminal cells in a subset of human prostate carcinomas. Star indicates histologically benign prostatic gland. Bar corresponds to 50 μm. See also Figure S3, Tables S4 and S5.