Main characteristics of study participants.

<p>PSA, prostate-specific antigen; CI, confidence interval; NA, not available.</p><p>Main characteristics of study participants.</p

Characterization of SEPT9_i1 antibodies and scoring of SEPT9_i1 staining intensity.

<p>HEK-293T embryonic human kidney cells were transiently transfected with Flag-SEPT9_i1 construct or empty vector (EV). (A) Whole cellular extracts were prepared and analyzed by 4–20% SDS-PAGE and immunoblotting (IB) with antibodies to Flag (1:2000), SEPT9_i1 (1:3000), preimmune serum (1:3000) or SEPT9_i1 antibody (1:3000) pre-incubated with 10 μM of the immunogen peptide for 4 hours. (B) The same cellular extracts were subjected to immunoprecipitation (IP) using anti-Flag antibody and the immuneprecipitates were subjected to 4–15% SDS-PAGE and then immunoblotted with anti-Flag or anti-SEPT9_i1 antibodies. (C) Representative SEPT9_i1 staining in human prostate cancer specimens. Score 0: no SEPT9_i1 staining, 1: low SEPT9_i1 staining 2: medium SEPT9_i1 staining and 3: high SEPT9_i1 staining. Magnification x200, scale bar 50 μm.</p

Gleason score correlation with SEPT9_i1 staining intensity.

<p>Mean SEPT9_i1 staining intensity ± SE of each Gleason score group (score 5: 3 patients, score 6: 8 patients, score 7: 19 patients, score 8: 3 patients, score 9: 6 patients and score 10: 2 patients) was calculated using either visual scoring (A) or automated image analysis with the ARIOL-SL50 system (B).</p

SEPT9_i1 staining in prostate cancer metastases.

<p>Metastatic prostate cancer lesions from bone marrow, lymph node and bone were immunostained with SEPT9_i1. Left panels are low (x100) magnification (LM) (scale bar 100 μm) and right panel are high (x200) magnification (HM) (scale bar 50 μm). Note high level of SEPT9_i1 staining in all metastases.</p

Correlation between "patients' characteristics" and SEPT9_i1 staining.

<p>PSA, prostate-specific antigen.</p><p>Correlation between "patients' characteristics" and SEPT9_i1 staining.</p

Supplementary figures 1-9 Supplementary Table 1 from Long Noncoding RNA MALAT1 Regulates Cancer Glucose Metabolism by Enhancing mTOR-Mediated Translation of TCF7L2

Supplementary Information: Figure S1: MALAT1 affects cancer glucose metabolism Figure S2: Glucose metabolism in HCC cell lines with MALAT1 knockdown. Figure S3: Regulation of TCF7L2 protein expression by MALAT1 in HCC cell lines. Figure S4: A non-phosphorylatable mutant of 4EBP1 inhibits TCF7L2 protein expression and expression of glycolytic genes. Figure S5: SRSF1 regulates TCF7L2 levels post-transcriptionally. Figure S6: TCF7L2 modulates glucose metabolism in a HCC cell line. Figure S7: MALAT1 and TCF7L2 regulate gluconeogenesis through the same pathway. Figure S8: Oncogenic properties of HCC cell lines with TCF7L2 knockdown. Figure S9: TCF7L2 protein, Gluconeogenesis and Glycolytic enzyme expression in livers from mouse HCC model Mdr2-/-. Table S1: List and sequences of shRNAs, siRNAs and PCR primers used in the paper</p

Immunohistochemical staining for COUP-TF1 in human prostate cancer, high grade PIN and benign epithelium.

<p>A. Immunohistochemical staining of human prostate cancer samples and adjacent benign glands for COUP-TF1 shows a nucleolar distribution of COUP-TF1 in malignant cells (upper left) and no COUP-TF1 staining in the adjacent benign gland (lower right). Stromal cells show nuclear staining of COUP-TF1. A representative of 28 samples analyzed is shown. B. Prostatic Intraepithelial Neoplasia (PIN) shows nucleolar distribution of COUP-TF1 and adjacent stromal cells show nuclear staining. C. immunostaining of LAPC4 xenografts shows nucleolar COUP-TF1 staining.</p

Genome-Wide Analysis of Androgen Receptor Targets Reveals COUP-TF1 as a Novel Player in Human Prostate Cancer

<div><p>Androgen activity plays a key role in prostate cancer progression. Androgen receptor (AR) is the main mediator of androgen activity in the prostate, through its ability to act as a transcription mediator. Here we performed a genome-wide analysis of human AR binding to promoters in the presence of an agonist or antagonist in an androgen dependent prostate cancer cell line. Many of the AR bound promoters are bound in all examined conditions while others are bound only in the presence of an agonist or antagonist. Several motifs are enriched in AR bound promoters, including the AR Response Element (ARE) half-site and recognition elements for the transcription factors OCT1 and SOX9. This suggests that these 3 factors could define a module of co-operating transcription factors in the prostate. Interestingly, AR bound promoters are preferentially located in AT rich genomic regions. Analysis of mRNA expression identified chicken ovalbumin upstream promoter-transcription factor 1 (COUP-TF1) as a direct AR target gene that is downregulated upon binding by the agonist liganded AR. COUP-TF1 immunostaining revealed nucleolar localization of COUP-TF1 in epithelium of human androgen dependent prostate cancer, but not in adjacent benign prostate epithelium. Stromal cells both in human and mouse prostate show nuclear COUP-TF1 staining. We further show that there is an inverse correlation between COUP-TF1 expression in prostate stromal cells and the rising levels of androgen with advancing puberty. This study extends the pool of recognized putative AR targets and identifies a negatively regulated target of AR – COUP-TF1 – which could possibly play a role in human prostate cancer.</p> </div

miRNA-Guided Imaging and Photodynamic Therapy Treatment of Cancer Cells Using Zn(II)-Protoporphyrin IX-Loaded Metal–Organic Framework Nanoparticles

An analytical platform for the selective miRNA-21-guided imaging of breast cancer cells and miRNA-221-guided imaging of ovarian cancer cells and the selective photodynamic therapy (PDT) of these cancer cells is introduced. The method is based on Zn­(II)-protoporphyrin IX, Zn­(II)-PPIX-loaded UiO-66 metal–organic framework nanoparticles, NMOFs, gated by two hairpins Hi/Hj through ligation of their phosphate residues to the vacant Zr4+-ions associated with the NMOFs. The hairpins are engineered to include the miRNA recognition sequence in the stem domain of Hi, and in the Hi and Hj, partial locked stem regions of G-quadruplex subunits. Intracellular phosphate-ions displace the hairpins, resulting in the release of the Zn­(II)-PPIX and intracellular miRNAs open Hi, and this triggers the autonomous cross-opening of Hi and Hj. This activates the interhairpin hybridization chain reaction and leads to the assembly of highly fluorescent Zn­(II)-PPIX-loaded G-quadruplex chains. The miRNA-guided fluorescent chains allow selective imaging of cancer cells. Moreover, PDT with visible light selectively kills cancer cells and tumor cells through the formation of toxic reactive oxygen species

Over represented Transfac motifs.

<p>Over represented Transfac motifs.</p