tcf7l2 Search Results


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  • 85
    Thermo Fisher tcf7l2 rs12255372
    Tcf7l2 Rs12255372, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher tcf7l2 rs10885409
    Tcf7l2 Rs10885409, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher tcf7l2 rs12255372 snp
    Tcf7l2 Rs12255372 Snp, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher tcf7l2 polymorphisms rs12255372
    (A and B) LD patterns between SNPs rs7901695, rs7903146, rs7895340, rs11196205, and <t>rs12255372</t> of <t>TCF7L2</t> in white (A) and black (B) ARIC participants. The LD pattern in A is similar to the one observed among the HapMap CEU sample. SNPs are labeled above
    Tcf7l2 Polymorphisms Rs12255372, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Abcam tcf7l2
    In in vitro HCV model up-regulates genes involved in lipid metabolism. Immunofluorescence assay HCV NS5A (red) (A) and lipid accumulation (green) (B). C) Real-time analysis (mean values± S.D) of ADIG, Adipsin, KLF15, KLF3, <t>TCF7L2</t> and TWIST 1 are expressed as fold change and normalized with a housekeeping gene. Asterisks indicate significant difference between J6/JFH1 HuH7.5 vs HuH7.5 (*p
    Tcf7l2, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 42 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher snp tcf7l2 c 29347861 10
    In in vitro HCV model up-regulates genes involved in lipid metabolism. Immunofluorescence assay HCV NS5A (red) (A) and lipid accumulation (green) (B). C) Real-time analysis (mean values± S.D) of ADIG, Adipsin, KLF15, KLF3, <t>TCF7L2</t> and TWIST 1 are expressed as fold change and normalized with a housekeeping gene. Asterisks indicate significant difference between J6/JFH1 HuH7.5 vs HuH7.5 (*p
    Snp Tcf7l2 C 29347861 10, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Abcam anti tcf7l2
    Increased expression of the Wnt effector transcription factor 7L2 <t>(TCF7L2)</t> in parthenogenetic morulae and blastocysts. Double immunostaining against TCF7L2 ( red ) and Cdx2 ( green ) revealed dramatically increased TCF7L2 expression (indicated by arrows )
    Anti Tcf7l2, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam tcf7l2 antibody
    EGLN2 negatively correlates with <t>TCF7L2</t> expression and is a prognostic indicator in pancreatic cancer patients. a EGLNs regulated hydroxylation of HIF-1α to ultimately affect HIF-1α stability. b – d The prognostic value of EGLNs family members, including EGLN1, EGLN2, and EGLN3, showed that decreased expression of EGLN2 predicted a worse prognosis for pancreatic cancer by analysis of the TCGA dataset. e – g Correlation analysis of the TCGA dataset indicated that TCF7L2 expression was negatively correlated with EGLN2, and was positively correlated with EGLN1 and EGLN3. h TCF7L2 knockdown resulted in an increase of EGLN2 levels in PANC-1 and MIA PaCa-2 cells. ** P
    Tcf7l2 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher tcf7l2
    Feeding stimulates <t>TCF7L2</t> expression. A : effect of fasting and refeeding (4 h) on hepatic TCF7L2 expression in C57BL/6 mice by Western blotting. Cyclin D1, a known downstream target of the Wnt signaling pathway. Representative blot ( n = 6/group). B and
    Tcf7l2, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 47 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore tcf7l2
    <t>TCF7L2</t> correlates with AR and FOXA1 in molecular apocrine breast cancers. ( A ) Endogenous TCF7L2 was coimmunoprecipitated with AR and FOXA1 in MDA-MB-453 breast cancer cells. Nuclear extracts (NEs) from vehicle (veh) or DHT-treated MDA-MB-435 cells were
    Tcf7l2, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 34 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam tcf7l2 ep20334
    <t>TCF7L2</t> correlates with AR and FOXA1 in molecular apocrine breast cancers. ( A ) Endogenous TCF7L2 was coimmunoprecipitated with AR and FOXA1 in MDA-MB-453 breast cancer cells. Nuclear extracts (NEs) from vehicle (veh) or DHT-treated MDA-MB-435 cells were
    Tcf7l2 Ep20334, supplied by Abcam, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc tcf7l2
    Association of <t>TCF7L2</t> and HNF4α in HepG2 cells . (a) HNF4α and FOXA2 ChIP-seq data were downloaded from the UCSC genome browser, and peaks were called and overlapped with the HepG2 cell type-specific TCF7L2 peaks. (b) Peaks bound only by HNF4α, only by TCF7L2, or by both factors were analyzed for the presence of HNF4α and TCF7L2 motifs. (c) For the set of 7,576 peaks bound by all three factors, the location of the HNF4α and FOXA2 peaks were plotted relative to the center of the TCF7L2 peak. (d) A comparison of TCF7L2, HNF4α, and FOXA2 binding patterns near the GREB1 locus is shown. The hg19 genomic coordinates are chr2:11,636,208-11,708,654. The number of tags reflecting the ChIP enrichments is plotted on the y-axis.
    Tcf7l2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 92/100, based on 200 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Merck KGaA dn tcf7l2
    Expression of <t>DN-TCF7L2</t> reduced SMK-17-induced apoptosis in HCT 116 cells. HCT 116 cells were transfected with DN-TCF7L2. (A) Luciferase activity was monitored through co-transfection with the TOPFlash and Renilla luciferase plasmids. (B) HCT 116 cells expressing DN-TCF7L2 were treated with the indicated concentrations of SMK-17 for 48 h and stained with annexin V/APC and 7-AAD. Cells were analyzed by flow cytometry with gates including only EGFP-positive cells.
    Dn Tcf7l2, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 89/100, based on 54 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Novus Biologicals tcf7l2
    A-C. Stained cells within the arcuate hypothalamic nucleus. A. NPY-stained cells showing immunoreactive fibers extending beyond the nucleus into the region adjacent to the arcuate nucleus. B. <t>TCF7L2</t> stained nuclei of arcuate neurons. C. Agouti-related
    Tcf7l2, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 91/100, based on 54 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher gene exp tcf7l2 hs01009038 m1
    Cumulative weight gain ( A ), absolute body weight ( B ), final body weight ( C ), and cumulative food intake ( D ) during and after 12 weeks of the HFD ( n = 14 <t>ΔE11-TCF7L2,</t> n = 16 LoxP controls). * P
    Gene Exp Tcf7l2 Hs01009038 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 87/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher gene exp tcf7l2 mm01261075 m1
    Characterization of <t>TCF7L2</t> up-regulation in response to 1,25(OH) 2 D 3 . ( A ) qPCR analysis of CaCo2 cells transfected and treated as described in Figure 2D and 2E . TCF7L2 (left panel) and CYP24A1 (right panel) transcripts. Data are normalized to GAPDH and plotted relative to each EtOH-treated control. p-values are derived from student's t-test comparison between EtOH and D 3 controls: * p
    Gene Exp Tcf7l2 Mm01261075 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology anti tcf7l2
    MUC1-C cytoplasmic domain associates with <t>TCF7L2.</t> A , lysates from ZR-75-1 ( left ) and MCF-7 ( right ) breast cancer cells were precipitated with anti-TCF7L2 or, as a control, non-immune IgG. The precipitates were immunoblotted with the indicated antibodies.
    Anti Tcf7l2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 88/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher gene exp tcf7l2 mm00501505 m1
    Cumulative weight gain ( A ), absolute body weight ( B ), final body weight ( C ), and cumulative food intake ( D ) during and after 12 weeks of the HFD ( n = 14 <t>ΔE11-TCF7L2,</t> n = 16 LoxP controls). * P
    Gene Exp Tcf7l2 Mm00501505 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti tcf7l2
    The TCF/LEF family of transcription factors bind the insertion allele of rs386772267 in an allele-specific manner. (A) Electromobility shift assay (EMSA) of rs386772267 deletion and insertion allele probes incubated with PANC-1 nuclear extract with and without unlabelled allelic competitors reveals an allele-specific band for the insertion probe (indicated by the arrow). (B) Alignment of binding sequence motifs for TCF/LEF transcription factors against the insertion probe (insertion allele designated in red). Similarity scores indicate similarity of aligned insertion allele probe to binding sequence matrices for the respective TFs as calculated by Genomatix MatInspector TM . (C and D) The ratios for medium and light labelled proteins bound to ( C ) insertion allele vs. scrambled insertion or ( D ) vs. deletion allele probes. Isotopic labeling for the y-axis experiments were reversed compared to those of the x-axis. Proteins indicated in red <t>(TCF7L2</t> and SNRPA1) showed consistent preferential binding to the insertion allele vs. both scrambled insertion and deletion allele probes. (E) The insertion probe’s allele-specific EMSA band (indicated by black arrow) demonstrated supershifting (indicated by red arrow) or disappeared entirely in the presence of LEF1, TCF7 or TCF7L2 antibodies using either MIAPaCa-2 or PANC-1 nuclear extract.
    Anti Tcf7l2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 88/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Thermo Fisher gene exp tcf7l2 hs00181036 m1
    The TCF/LEF family of transcription factors bind the insertion allele of rs386772267 in an allele-specific manner. (A) Electromobility shift assay (EMSA) of rs386772267 deletion and insertion allele probes incubated with PANC-1 nuclear extract with and without unlabelled allelic competitors reveals an allele-specific band for the insertion probe (indicated by the arrow). (B) Alignment of binding sequence motifs for TCF/LEF transcription factors against the insertion probe (insertion allele designated in red). Similarity scores indicate similarity of aligned insertion allele probe to binding sequence matrices for the respective TFs as calculated by Genomatix MatInspector TM . (C and D) The ratios for medium and light labelled proteins bound to ( C ) insertion allele vs. scrambled insertion or ( D ) vs. deletion allele probes. Isotopic labeling for the y-axis experiments were reversed compared to those of the x-axis. Proteins indicated in red <t>(TCF7L2</t> and SNRPA1) showed consistent preferential binding to the insertion allele vs. both scrambled insertion and deletion allele probes. (E) The insertion probe’s allele-specific EMSA band (indicated by black arrow) demonstrated supershifting (indicated by red arrow) or disappeared entirely in the presence of LEF1, TCF7 or TCF7L2 antibodies using either MIAPaCa-2 or PANC-1 nuclear extract.
    Gene Exp Tcf7l2 Hs00181036 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Santa Cruz Biotechnology tcf7l2
    A – E , Response to mechanical injury in <t>TCF7L2</t> (transcription factor 7-like 2) overexpressing mice. A , Immunofluorescence (IF) staining of TCF7L2 in wild-type (WT) mouse carotid at baseline and post guidewire injury (relative fluorescent intensities/area and the tunica media (M) area are shown next to the figure. B , TCF7L2 IF intensity in heterozygote overexpressing (TCF7L2-bac) and WT mice before and after injury (relative fluorescent intensities shown underneath). C , Neointima (NI) formation and EVG (elastic tissue fibers-Verhoeff’s Van Gieson) staining in carotid arteries of WT and TCF7L2-bac mice postguidewire injury. Quantifications of NI for both female and male WT and TCF7L2-bac mice postcarotid guidewire injury are shown underneath corresponding figures. The quantification is done by the ratio of intima/M surface area. D and E , Immunofluorescent assessment of SM-MHC (smooth muscle cell myosin heavy chain) and α-SMA (alpha smooth muscle cell actin; both in green) in carotids of TCF7L2-bac, and WT mice, before and after guidewire injury. The relative fluorescent intensities shown underneath. The yellow dotted lined in ( A ) demarcate the NI. *, **, ***, ****Significance with P
    Tcf7l2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 181 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Proteintech tcf7l2
    Adiponectin inhibited the expression of β-catenin-associated transcription factor <t>TCF7L2</t> in pancreatic cancer cells. A and B, The pooled RNA sample from the adiponectin-treated BxPC-3 cells or control cells was subjected to microarray analysis of the mRNA expression profile. A total of 180 genes were identified to be differentially expressed at the mRNA level in response to adiponectin treatment. A, Hierarchical clustering of the 180 differentially expressed genes. The colored images are presented as described; the green and red colors indicate low and high expression levels, respectively. B, GO annotation of differentially expressed genes in BxPC-3 cells induced by adiponectin treatment. The overrepresented GO terms ( P
    Tcf7l2, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore human tcf7l2
    Distribution of LEF1, <t>TCF7L2,</t> and β-catenin proteins in the adult prethalamus and epithalamus. a TCF7L2 and nuclear β-catenin are present in the subsets of cells in the pregeniculate nucleus. b A clear border is seen between LEF1-, TCF7L2-, and nuclear β-catenin-positive cells between the thalamus and reticular thalamic nucleus. c In the habenula, LEF1, TCF7L2, and nuclear β-catenin are present only in the medial part. DLG dorsal lateral geniculate; LHb lateral habenula; MHb medial habenula; PG pregeniculate nucleus; RT reticular thalamic nucleus; VPL ventral posterolateral thalamic nucleus. Scale bar 100 μm
    Human Tcf7l2, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Upstate Biotechnology Inc anti tcf7l2
    <t>TCF7L2</t> protein expression . Western Blot showing lower amounts of TCF7L2 protein expression in MDA-MB-231 EpCAM cells compared to their empty vector counterparts.
    Anti Tcf7l2, supplied by Upstate Biotechnology Inc, used in various techniques. Bioz Stars score: 88/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Genoway tcf7l2
    Generation of pancreas-specific <t>Tcf7l2</t> null mice. a) Schematic showing wild-type (top) and conditional inactivation (bottom) of the Tcf7l2 allelle with binding sites for primers used for genotyping (dotted lines). Black boxes represent exons with exon number indicated in white. b) Immunohistochemical analysis of pancreata for Tcf7l2 and insulin. Scale bar = 20 micrometre. c) Real time PCR analysis of gene expression in islets of pTcf7l2 mice (black bar) and wild-type littermate control (dotted/grey bar). Tcf7l2 gene expression was undetected (UD; Ct > 40) in pTcf7l2 mice. Ccnd1, cyclin D1 gene.
    Tcf7l2, supplied by Genoway, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    (A and B) LD patterns between SNPs rs7901695, rs7903146, rs7895340, rs11196205, and rs12255372 of TCF7L2 in white (A) and black (B) ARIC participants. The LD pattern in A is similar to the one observed among the HapMap CEU sample. SNPs are labeled above

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: TCF7L2 Variants Associate with CKD Progression and Renal Function in Population-Based Cohorts

    doi: 10.1681/ASN.2007121291

    Figure Lengend Snippet: (A and B) LD patterns between SNPs rs7901695, rs7903146, rs7895340, rs11196205, and rs12255372 of TCF7L2 in white (A) and black (B) ARIC participants. The LD pattern in A is similar to the one observed among the HapMap CEU sample. SNPs are labeled above

    Article Snippet: Genotyping of TCF7L2 polymorphisms rs12255372, rs7903146, rs7901695, rs11196205, and rs7895340 in ARIC was performed separately using the TaqMan assay (Applied Biosystems, Foster City, CA) by the ARIC Central DNA Laboratory.

    Techniques: Labeling

    In in vitro HCV model up-regulates genes involved in lipid metabolism. Immunofluorescence assay HCV NS5A (red) (A) and lipid accumulation (green) (B). C) Real-time analysis (mean values± S.D) of ADIG, Adipsin, KLF15, KLF3, TCF7L2 and TWIST 1 are expressed as fold change and normalized with a housekeeping gene. Asterisks indicate significant difference between J6/JFH1 HuH7.5 vs HuH7.5 (*p

    Journal: PLoS ONE

    Article Title: Lymphocytes as Liver Damage Mirror of HCV Related Adipogenesis Deregulation

    doi: 10.1371/journal.pone.0092343

    Figure Lengend Snippet: In in vitro HCV model up-regulates genes involved in lipid metabolism. Immunofluorescence assay HCV NS5A (red) (A) and lipid accumulation (green) (B). C) Real-time analysis (mean values± S.D) of ADIG, Adipsin, KLF15, KLF3, TCF7L2 and TWIST 1 are expressed as fold change and normalized with a housekeeping gene. Asterisks indicate significant difference between J6/JFH1 HuH7.5 vs HuH7.5 (*p

    Article Snippet: HCV up-regulated genes are closely related to virological and biochemical liver damage parameters, as well as to the severity of steatosis and fibrosis Statistical analysis statistically positively correlated the HCV viremia plasma levels (VL) and antibody titer (AbT) to Adipogenin (ADIG), Adipsin (CFD), KLF15, KLF3, TCF7L2 and Twist1, either in HCV+ PBMCs or in liver tissues ( ,*p < 0.05).

    Techniques: In Vitro, Immunofluorescence

    HCV up-regulates genes involved in lipid metabolism but with addictional role in inflammation, proliferation and transformation. Real-time analysis (mean values± S.D) of ADIG and Adipsin (A), KLF15 and KLF3 (B), TCF7L2 and TWIST 1 (C) are expressed as fold change, and normalized with an housekeeping gene. Asterisks indicate significant difference between HCV+ vs HDs (*p

    Journal: PLoS ONE

    Article Title: Lymphocytes as Liver Damage Mirror of HCV Related Adipogenesis Deregulation

    doi: 10.1371/journal.pone.0092343

    Figure Lengend Snippet: HCV up-regulates genes involved in lipid metabolism but with addictional role in inflammation, proliferation and transformation. Real-time analysis (mean values± S.D) of ADIG and Adipsin (A), KLF15 and KLF3 (B), TCF7L2 and TWIST 1 (C) are expressed as fold change, and normalized with an housekeeping gene. Asterisks indicate significant difference between HCV+ vs HDs (*p

    Article Snippet: HCV up-regulated genes are closely related to virological and biochemical liver damage parameters, as well as to the severity of steatosis and fibrosis Statistical analysis statistically positively correlated the HCV viremia plasma levels (VL) and antibody titer (AbT) to Adipogenin (ADIG), Adipsin (CFD), KLF15, KLF3, TCF7L2 and Twist1, either in HCV+ PBMCs or in liver tissues ( ,*p < 0.05).

    Techniques: Transformation Assay

    Increased expression of the Wnt effector transcription factor 7L2 (TCF7L2) in parthenogenetic morulae and blastocysts. Double immunostaining against TCF7L2 ( red ) and Cdx2 ( green ) revealed dramatically increased TCF7L2 expression (indicated by arrows )

    Journal: Stem Cells and Development

    Article Title: Epigenetic Disruptions of Histone Signatures for the Trophectoderm and Inner Cell Mass in Mouse Parthenogenetic Embryos

    doi: 10.1089/scd.2014.0310

    Figure Lengend Snippet: Increased expression of the Wnt effector transcription factor 7L2 (TCF7L2) in parthenogenetic morulae and blastocysts. Double immunostaining against TCF7L2 ( red ) and Cdx2 ( green ) revealed dramatically increased TCF7L2 expression (indicated by arrows )

    Article Snippet: After blocking, embryos were incubated at 4°C overnight with the following primary antibodies diluted in blocking solution: anti-acetylated histone H3 lysine 9 (rabbit polyclonal, 1:200 dilution; Abcam), anti-CARM1 (both mouse monoclonal and rabbit polyclonal, 1:200 dilution; Abcam), anti-Cdx2 (mouse monoclonal, 1:100 dilution; BioGenex), anti-dimethylated histone H3 arginine 26 (rabbit polyclonal, 1:200 dilution; Abcam), anti-Ezh2 (rabbit polyclonal, 1:200 dilution; Millipore), anti-mono-ubiquitinated histone H2A lysine 119 (mouse monoclonal, 1:200 dilution; Millipore), anti-phosphorylated Akt1 (phospho-S473) (rabbit polyclonal, 1:100 dilution; Abcam), anti-phosphorylated Ezh2 (phospho-S21) (rabbit polyclonal, 1:100 dilution; Abcam), anti-TCF7L2 (rabbit polyclonal, 1:100 dilution; Abcam), anti-trimethylated histone H3 lysine 4 (rabbit polyclonal, 1:200 dilution; Abcam), anti-trimethylated histone H3 lysine 9 (rabbit polyclonal, 1:200 dilution; Millipore), or anti-trimethylated histone H3 lysine 27 (rabbit polyclonal, 1:200 dilution; Millipore).

    Techniques: Expressing, Double Immunostaining

    EGLN2 negatively correlates with TCF7L2 expression and is a prognostic indicator in pancreatic cancer patients. a EGLNs regulated hydroxylation of HIF-1α to ultimately affect HIF-1α stability. b – d The prognostic value of EGLNs family members, including EGLN1, EGLN2, and EGLN3, showed that decreased expression of EGLN2 predicted a worse prognosis for pancreatic cancer by analysis of the TCGA dataset. e – g Correlation analysis of the TCGA dataset indicated that TCF7L2 expression was negatively correlated with EGLN2, and was positively correlated with EGLN1 and EGLN3. h TCF7L2 knockdown resulted in an increase of EGLN2 levels in PANC-1 and MIA PaCa-2 cells. ** P

    Journal: Cell Death & Disease

    Article Title: TCF7L2 positively regulates aerobic glycolysis via the EGLN2/HIF-1α axis and indicates prognosis in pancreatic cancer

    doi: 10.1038/s41419-018-0367-6

    Figure Lengend Snippet: EGLN2 negatively correlates with TCF7L2 expression and is a prognostic indicator in pancreatic cancer patients. a EGLNs regulated hydroxylation of HIF-1α to ultimately affect HIF-1α stability. b – d The prognostic value of EGLNs family members, including EGLN1, EGLN2, and EGLN3, showed that decreased expression of EGLN2 predicted a worse prognosis for pancreatic cancer by analysis of the TCGA dataset. e – g Correlation analysis of the TCGA dataset indicated that TCF7L2 expression was negatively correlated with EGLN2, and was positively correlated with EGLN1 and EGLN3. h TCF7L2 knockdown resulted in an increase of EGLN2 levels in PANC-1 and MIA PaCa-2 cells. ** P

    Article Snippet: TCF7L2 antibody (Abcam, ab76151) was used at a dilution of 1:100.

    Techniques: Expressing

    TCF7L2 targets EGLN2 to regulate the HIF-1α axis in pancreatic cancer. a Schematic representation of the ELGN2 promoter regions has shown that EGLN2 promoter region contains putative TCF7L2-binding elements. b We cloned the promoter region of EGLN2, ranging from −3000 to +200, into pGL3-Basic vector, and observed that TCF7L2 inhibited EGLN2 promoter activity. c Dual luciferase assay results suggested that TCF7L2 could suppress EGLN2 promoter activity (** P

    Journal: Cell Death & Disease

    Article Title: TCF7L2 positively regulates aerobic glycolysis via the EGLN2/HIF-1α axis and indicates prognosis in pancreatic cancer

    doi: 10.1038/s41419-018-0367-6

    Figure Lengend Snippet: TCF7L2 targets EGLN2 to regulate the HIF-1α axis in pancreatic cancer. a Schematic representation of the ELGN2 promoter regions has shown that EGLN2 promoter region contains putative TCF7L2-binding elements. b We cloned the promoter region of EGLN2, ranging from −3000 to +200, into pGL3-Basic vector, and observed that TCF7L2 inhibited EGLN2 promoter activity. c Dual luciferase assay results suggested that TCF7L2 could suppress EGLN2 promoter activity (** P

    Article Snippet: TCF7L2 antibody (Abcam, ab76151) was used at a dilution of 1:100.

    Techniques: Binding Assay, Clone Assay, Plasmid Preparation, Activity Assay, Luciferase

    TCF7L2 positively regulates the proliferation of pancreatic cancer. a Quantitative RT-PCR analysis of TCF7L2 knockdown efficiency in PANC-1 and MIA PaCa-2 cell lines that stably express shRNA oligoes against TCF7L2. ** P

    Journal: Cell Death & Disease

    Article Title: TCF7L2 positively regulates aerobic glycolysis via the EGLN2/HIF-1α axis and indicates prognosis in pancreatic cancer

    doi: 10.1038/s41419-018-0367-6

    Figure Lengend Snippet: TCF7L2 positively regulates the proliferation of pancreatic cancer. a Quantitative RT-PCR analysis of TCF7L2 knockdown efficiency in PANC-1 and MIA PaCa-2 cell lines that stably express shRNA oligoes against TCF7L2. ** P

    Article Snippet: TCF7L2 antibody (Abcam, ab76151) was used at a dilution of 1:100.

    Techniques: Quantitative RT-PCR, Stable Transfection, shRNA

    TCF7L2 is positively correlated with HIF-1α stability and relevant glycolysis genes (GLUT1, HK2, LDHA) in pancreatic cancer. a TCF7L2 silencing decreased the protein level of HIF-1α. b There was no significant change in the expression of HIF-1α at the transcriptional level in TCF7L2-silenced PANC-1 and MIA PaCa-2 cells. c – f Kinetics of HIF-1α protein degradation. HIF-1α protein degraded faster in shTCF7L2 cells than in controls in PANC-1 ( c , e ) and MIA PaCa-2 cells ( d , f ). g TCF7L2 could upregulate HRE-luciferase activity in HEK293T cells. ** P

    Journal: Cell Death & Disease

    Article Title: TCF7L2 positively regulates aerobic glycolysis via the EGLN2/HIF-1α axis and indicates prognosis in pancreatic cancer

    doi: 10.1038/s41419-018-0367-6

    Figure Lengend Snippet: TCF7L2 is positively correlated with HIF-1α stability and relevant glycolysis genes (GLUT1, HK2, LDHA) in pancreatic cancer. a TCF7L2 silencing decreased the protein level of HIF-1α. b There was no significant change in the expression of HIF-1α at the transcriptional level in TCF7L2-silenced PANC-1 and MIA PaCa-2 cells. c – f Kinetics of HIF-1α protein degradation. HIF-1α protein degraded faster in shTCF7L2 cells than in controls in PANC-1 ( c , e ) and MIA PaCa-2 cells ( d , f ). g TCF7L2 could upregulate HRE-luciferase activity in HEK293T cells. ** P

    Article Snippet: TCF7L2 antibody (Abcam, ab76151) was used at a dilution of 1:100.

    Techniques: Expressing, Luciferase, Activity Assay

    TCF7L2 is positively related with aerobic glycolysis and 18 F-FDG uptake in pancreatic cancer. a , b TCF7L2 knockdown decreased glycolysis capacity, which could be shown by the ECAR test in PANC-1 and MIA PaCa-2 cells. c , d OCR significantly increased in TCF7L2-silenced PANC-1 and MIA PaCa-2 cells, indicating TCF7L2 functions as a positive regulator of mitochondrial respiration. e , f TCF7L2 expression positively correlated with 18 F-FDG uptake in the in vivo xenograft model. ** P

    Journal: Cell Death & Disease

    Article Title: TCF7L2 positively regulates aerobic glycolysis via the EGLN2/HIF-1α axis and indicates prognosis in pancreatic cancer

    doi: 10.1038/s41419-018-0367-6

    Figure Lengend Snippet: TCF7L2 is positively related with aerobic glycolysis and 18 F-FDG uptake in pancreatic cancer. a , b TCF7L2 knockdown decreased glycolysis capacity, which could be shown by the ECAR test in PANC-1 and MIA PaCa-2 cells. c , d OCR significantly increased in TCF7L2-silenced PANC-1 and MIA PaCa-2 cells, indicating TCF7L2 functions as a positive regulator of mitochondrial respiration. e , f TCF7L2 expression positively correlated with 18 F-FDG uptake in the in vivo xenograft model. ** P

    Article Snippet: TCF7L2 antibody (Abcam, ab76151) was used at a dilution of 1:100.

    Techniques: Expressing, In Vivo

    Feeding stimulates TCF7L2 expression. A : effect of fasting and refeeding (4 h) on hepatic TCF7L2 expression in C57BL/6 mice by Western blotting. Cyclin D1, a known downstream target of the Wnt signaling pathway. Representative blot ( n = 6/group). B and

    Journal: American Journal of Physiology - Endocrinology and Metabolism

    Article Title: The Wnt signaling pathway effector TCF7L2 is upregulated by insulin and represses hepatic gluconeogenesis

    doi: 10.1152/ajpendo.00249.2012

    Figure Lengend Snippet: Feeding stimulates TCF7L2 expression. A : effect of fasting and refeeding (4 h) on hepatic TCF7L2 expression in C57BL/6 mice by Western blotting. Cyclin D1, a known downstream target of the Wnt signaling pathway. Representative blot ( n = 6/group). B and

    Article Snippet: Transfection of 5 nM small interfering RNA (siRNA) recognizing either a scrambled sequence or TCF7L2 (Ambion) was achieved using Lipofectamine RNAiMAX (Invitrogen) per the manufacturer's instructions.

    Techniques: Expressing, Mouse Assay, Western Blot

    A schematic presentation of the role of Wnt signaling and TCF7L2 in hepatic gluconeogenesis. Feeding upregulates plasma insulin levels, which leads to β-cat Ser 675 phosphorylation and TCF7L2 expression. A : increased levels of TCF7L2 may repress

    Journal: American Journal of Physiology - Endocrinology and Metabolism

    Article Title: The Wnt signaling pathway effector TCF7L2 is upregulated by insulin and represses hepatic gluconeogenesis

    doi: 10.1152/ajpendo.00249.2012

    Figure Lengend Snippet: A schematic presentation of the role of Wnt signaling and TCF7L2 in hepatic gluconeogenesis. Feeding upregulates plasma insulin levels, which leads to β-cat Ser 675 phosphorylation and TCF7L2 expression. A : increased levels of TCF7L2 may repress

    Article Snippet: Transfection of 5 nM small interfering RNA (siRNA) recognizing either a scrambled sequence or TCF7L2 (Ambion) was achieved using Lipofectamine RNAiMAX (Invitrogen) per the manufacturer's instructions.

    Techniques: Expressing

    Active Wnt signaling and transcription factor 7-like 2 (TCF7L2) expression are present in hepatocytes. A , top : depiction of the TOPGAL mouse transgene. A , bottom : β-galactosidase (LacZ) staining showing active Wnt activity in pericentral hepatocytes

    Journal: American Journal of Physiology - Endocrinology and Metabolism

    Article Title: The Wnt signaling pathway effector TCF7L2 is upregulated by insulin and represses hepatic gluconeogenesis

    doi: 10.1152/ajpendo.00249.2012

    Figure Lengend Snippet: Active Wnt signaling and transcription factor 7-like 2 (TCF7L2) expression are present in hepatocytes. A , top : depiction of the TOPGAL mouse transgene. A , bottom : β-galactosidase (LacZ) staining showing active Wnt activity in pericentral hepatocytes

    Article Snippet: Transfection of 5 nM small interfering RNA (siRNA) recognizing either a scrambled sequence or TCF7L2 (Ambion) was achieved using Lipofectamine RNAiMAX (Invitrogen) per the manufacturer's instructions.

    Techniques: Expressing, Staining, Activity Assay

    Insulin stimulates TCF7L2 expression in 2 hepatic cell lines. Effect of 100 nM insulin (4 h) on TCF7L2 expression assessed by Western blotting ( A and B ) and quantitative RT-PCR ( C and D ) in HepG2 and Hepa1–6 cell lines. E and F : effect of 100

    Journal: American Journal of Physiology - Endocrinology and Metabolism

    Article Title: The Wnt signaling pathway effector TCF7L2 is upregulated by insulin and represses hepatic gluconeogenesis

    doi: 10.1152/ajpendo.00249.2012

    Figure Lengend Snippet: Insulin stimulates TCF7L2 expression in 2 hepatic cell lines. Effect of 100 nM insulin (4 h) on TCF7L2 expression assessed by Western blotting ( A and B ) and quantitative RT-PCR ( C and D ) in HepG2 and Hepa1–6 cell lines. E and F : effect of 100

    Article Snippet: Transfection of 5 nM small interfering RNA (siRNA) recognizing either a scrambled sequence or TCF7L2 (Ambion) was achieved using Lipofectamine RNAiMAX (Invitrogen) per the manufacturer's instructions.

    Techniques: Expressing, Western Blot, Quantitative RT-PCR

    Knockdown of TCF7L2 upregulates hepatic gluconeogenesis. A : RNA sequences of TCF7L2 small interfering RNA (siRNA) oligonucleotides utilized in this study. The human TCF7L2 sequence is recognized by siRNA-s74839, whereas the mouse sequence is recognized

    Journal: American Journal of Physiology - Endocrinology and Metabolism

    Article Title: The Wnt signaling pathway effector TCF7L2 is upregulated by insulin and represses hepatic gluconeogenesis

    doi: 10.1152/ajpendo.00249.2012

    Figure Lengend Snippet: Knockdown of TCF7L2 upregulates hepatic gluconeogenesis. A : RNA sequences of TCF7L2 small interfering RNA (siRNA) oligonucleotides utilized in this study. The human TCF7L2 sequence is recognized by siRNA-s74839, whereas the mouse sequence is recognized

    Article Snippet: Transfection of 5 nM small interfering RNA (siRNA) recognizing either a scrambled sequence or TCF7L2 (Ambion) was achieved using Lipofectamine RNAiMAX (Invitrogen) per the manufacturer's instructions.

    Techniques: Small Interfering RNA, Sequencing

    TCF7L2 correlates with AR and FOXA1 in molecular apocrine breast cancers. ( A ) Endogenous TCF7L2 was coimmunoprecipitated with AR and FOXA1 in MDA-MB-453 breast cancer cells. Nuclear extracts (NEs) from vehicle (veh) or DHT-treated MDA-MB-435 cells were

    Journal: Genes & Development

    Article Title: Amplitude modulation of androgen signaling by c-MYC

    doi: 10.1101/gad.209569.112

    Figure Lengend Snippet: TCF7L2 correlates with AR and FOXA1 in molecular apocrine breast cancers. ( A ) Endogenous TCF7L2 was coimmunoprecipitated with AR and FOXA1 in MDA-MB-453 breast cancer cells. Nuclear extracts (NEs) from vehicle (veh) or DHT-treated MDA-MB-435 cells were

    Article Snippet: Western blot analysis was performed as described previously ( ) using antibodies against AR (sc-7305), MYC (sc-764), MAD1 (sc-222), MAX (sc-765), HER3 (sc-7390), TBP (sc-204), and GAPDH (sc-25778) from Santa Cruz Biotechnology; TCF7L2 (clone 1B1) from Sigma-Aldrich; FOXA1 (ab40868) from Abcam; and p-HER3 (no.4791), p-AKT (no.4060), and AKT (no.9272) from Cell Signaling.

    Techniques: Multiple Displacement Amplification

    Cooperation of AR, FOXA1, and TCF7L2 regulates the transcriptional activation of MYC . ( A ) Schematic graph shows the AR–FOXA1–TCF7L2 sites (shaded bars) within the MYC gene locus as defined by ChIP-seq in MDA-MB-453 cells. ( B ) 3C analysis

    Journal: Genes & Development

    Article Title: Amplitude modulation of androgen signaling by c-MYC

    doi: 10.1101/gad.209569.112

    Figure Lengend Snippet: Cooperation of AR, FOXA1, and TCF7L2 regulates the transcriptional activation of MYC . ( A ) Schematic graph shows the AR–FOXA1–TCF7L2 sites (shaded bars) within the MYC gene locus as defined by ChIP-seq in MDA-MB-453 cells. ( B ) 3C analysis

    Article Snippet: Western blot analysis was performed as described previously ( ) using antibodies against AR (sc-7305), MYC (sc-764), MAD1 (sc-222), MAX (sc-765), HER3 (sc-7390), TBP (sc-204), and GAPDH (sc-25778) from Santa Cruz Biotechnology; TCF7L2 (clone 1B1) from Sigma-Aldrich; FOXA1 (ab40868) from Abcam; and p-HER3 (no.4791), p-AKT (no.4060), and AKT (no.9272) from Cell Signaling.

    Techniques: Activation Assay, Chromatin Immunoprecipitation, Multiple Displacement Amplification

    TCF7L2 collaborates with AR and FOXA1 in the control of DHT-mediated transcriptional activation. ( A , B ) Correlation between DHT-up-regulated gene expression and the binding sites of AR, FOXA1, and TCF7L2 in all possible combinations. ( A ) The fold enrichment

    Journal: Genes & Development

    Article Title: Amplitude modulation of androgen signaling by c-MYC

    doi: 10.1101/gad.209569.112

    Figure Lengend Snippet: TCF7L2 collaborates with AR and FOXA1 in the control of DHT-mediated transcriptional activation. ( A , B ) Correlation between DHT-up-regulated gene expression and the binding sites of AR, FOXA1, and TCF7L2 in all possible combinations. ( A ) The fold enrichment

    Article Snippet: Western blot analysis was performed as described previously ( ) using antibodies against AR (sc-7305), MYC (sc-764), MAD1 (sc-222), MAX (sc-765), HER3 (sc-7390), TBP (sc-204), and GAPDH (sc-25778) from Santa Cruz Biotechnology; TCF7L2 (clone 1B1) from Sigma-Aldrich; FOXA1 (ab40868) from Abcam; and p-HER3 (no.4791), p-AKT (no.4060), and AKT (no.9272) from Cell Signaling.

    Techniques: Activation Assay, Expressing, Binding Assay

    Association of TCF7L2 and HNF4α in HepG2 cells . (a) HNF4α and FOXA2 ChIP-seq data were downloaded from the UCSC genome browser, and peaks were called and overlapped with the HepG2 cell type-specific TCF7L2 peaks. (b) Peaks bound only by HNF4α, only by TCF7L2, or by both factors were analyzed for the presence of HNF4α and TCF7L2 motifs. (c) For the set of 7,576 peaks bound by all three factors, the location of the HNF4α and FOXA2 peaks were plotted relative to the center of the TCF7L2 peak. (d) A comparison of TCF7L2, HNF4α, and FOXA2 binding patterns near the GREB1 locus is shown. The hg19 genomic coordinates are chr2:11,636,208-11,708,654. The number of tags reflecting the ChIP enrichments is plotted on the y-axis.

    Journal: Genome Biology

    Article Title: Cell type-specific binding patterns reveal that TCF7L2 can be tethered to the genome by association with GATA3

    doi: 10.1186/gb-2012-13-9-r52

    Figure Lengend Snippet: Association of TCF7L2 and HNF4α in HepG2 cells . (a) HNF4α and FOXA2 ChIP-seq data were downloaded from the UCSC genome browser, and peaks were called and overlapped with the HepG2 cell type-specific TCF7L2 peaks. (b) Peaks bound only by HNF4α, only by TCF7L2, or by both factors were analyzed for the presence of HNF4α and TCF7L2 motifs. (c) For the set of 7,576 peaks bound by all three factors, the location of the HNF4α and FOXA2 peaks were plotted relative to the center of the TCF7L2 peak. (d) A comparison of TCF7L2, HNF4α, and FOXA2 binding patterns near the GREB1 locus is shown. The hg19 genomic coordinates are chr2:11,636,208-11,708,654. The number of tags reflecting the ChIP enrichments is plotted on the y-axis.

    Article Snippet: To determine if we had identified the majority of the TCF7L2 peaks in each cell type, we performed a saturation analysis.

    Techniques: Chromatin Immunoprecipitation, Binding Assay

    Two modes of TCF7L2-mediated transcriptional repression of GATA3 target genes . (a) GATA3 tethers TCF7L2 to the genome and both factors cooperate to repress target genes. (b) GATA3 tethers TCF7L2 to the genome with TCF7L2 antagonizing GATA3-mediated transcriptional activation.

    Journal: Genome Biology

    Article Title: Cell type-specific binding patterns reveal that TCF7L2 can be tethered to the genome by association with GATA3

    doi: 10.1186/gb-2012-13-9-r52

    Figure Lengend Snippet: Two modes of TCF7L2-mediated transcriptional repression of GATA3 target genes . (a) GATA3 tethers TCF7L2 to the genome and both factors cooperate to repress target genes. (b) GATA3 tethers TCF7L2 to the genome with TCF7L2 antagonizing GATA3-mediated transcriptional activation.

    Article Snippet: To determine if we had identified the majority of the TCF7L2 peaks in each cell type, we performed a saturation analysis.

    Techniques: Activation Assay

    Transcriptional regulation of TCF7L2 and GATA3 target genes . (a) The nearest gene to each TCF7L2 binding site and the nearest gene to each GATA3 binding site was identified and the two lists were compared to identify 3,614 genes that are potentially regulated by both GATA3 and TCF7L2. (b) The expression of the 3,614 GATA3+TCF7L2 bound genes was analyzed in control cells, cells treated with siRNAs to TCF7L2, and cells treated with siRNAs to GATA3; the number of genes whose expression increases or decreases is shown. (c) A scatterplot of expression data from RNA-seq experiments. Each point corresponds to one NCBI Reference Sequence (RefSeq) transcript with fragments per kilobase of gene per million reads (FPKM) values for control and siGATA3 or control and siTCF7L2 knockdown samples shown on a log10 scale. The dashed line represents no change in gene expression between the two samples. Differentially expressed genes whose function corresponds to Gene Ontology categories of breast cancer, cell differentiation, and response to hormone stimulus are highlighted.

    Journal: Genome Biology

    Article Title: Cell type-specific binding patterns reveal that TCF7L2 can be tethered to the genome by association with GATA3

    doi: 10.1186/gb-2012-13-9-r52

    Figure Lengend Snippet: Transcriptional regulation of TCF7L2 and GATA3 target genes . (a) The nearest gene to each TCF7L2 binding site and the nearest gene to each GATA3 binding site was identified and the two lists were compared to identify 3,614 genes that are potentially regulated by both GATA3 and TCF7L2. (b) The expression of the 3,614 GATA3+TCF7L2 bound genes was analyzed in control cells, cells treated with siRNAs to TCF7L2, and cells treated with siRNAs to GATA3; the number of genes whose expression increases or decreases is shown. (c) A scatterplot of expression data from RNA-seq experiments. Each point corresponds to one NCBI Reference Sequence (RefSeq) transcript with fragments per kilobase of gene per million reads (FPKM) values for control and siGATA3 or control and siTCF7L2 knockdown samples shown on a log10 scale. The dashed line represents no change in gene expression between the two samples. Differentially expressed genes whose function corresponds to Gene Ontology categories of breast cancer, cell differentiation, and response to hormone stimulus are highlighted.

    Article Snippet: To determine if we had identified the majority of the TCF7L2 peaks in each cell type, we performed a saturation analysis.

    Techniques: Binding Assay, Expressing, RNA Sequencing Assay, Sequencing, Cell Differentiation

    Cell type-specific binding of TCF7L2 . (a,b) The ChIP-seq binding patterns of TCF7L2 are compared in six cell lines, demonstrating both common peaks (a) and cell type-specific binding (b). (c) The ChIP-seq binding patterns of TCF7L2 near and within the SH3BP4 locus is shown for three cell lines. The number of tags reflecting the ChIP enrichments are plotted on the y-axis; the chromosomal coordinates (hg19) shown are: (a) chr19:7,701,591-7,718,750; (b) chr1:112,997,195-113,019,766; and (c) chr2:235,767,270-235,974,731.

    Journal: Genome Biology

    Article Title: Cell type-specific binding patterns reveal that TCF7L2 can be tethered to the genome by association with GATA3

    doi: 10.1186/gb-2012-13-9-r52

    Figure Lengend Snippet: Cell type-specific binding of TCF7L2 . (a,b) The ChIP-seq binding patterns of TCF7L2 are compared in six cell lines, demonstrating both common peaks (a) and cell type-specific binding (b). (c) The ChIP-seq binding patterns of TCF7L2 near and within the SH3BP4 locus is shown for three cell lines. The number of tags reflecting the ChIP enrichments are plotted on the y-axis; the chromosomal coordinates (hg19) shown are: (a) chr19:7,701,591-7,718,750; (b) chr1:112,997,195-113,019,766; and (c) chr2:235,767,270-235,974,731.

    Article Snippet: To determine if we had identified the majority of the TCF7L2 peaks in each cell type, we performed a saturation analysis.

    Techniques: Binding Assay, Chromatin Immunoprecipitation

    TCF7L2 binding sites are distal and enriched for active enhancer histone marks . (a) Shown for the TCF7L2 binding sites in the six cell types and for the 1,864 peaks commonly bound in all six cells is the percentage of TCF7L2 binding sites in different genomic regions (hg19) relative to the nearest transcription start site (TSS). (b) The percentage of active enhancer regions containing a TCF7L2 binding site; active enhancers were defined by taking the regions that have an overlap of H3K4me1 and H3K27ac ChIP-seq peaks for the given cell line. (c) Heatmaps of the ChIP-Seq tags for H3K27ac and H3K4me1 at TCF7L2-bound regions (±3 kb windows around the center of all TCF7L2 peaks) for each cell line were generated by k-means cluster analysis. (d) The average RNA polymerase II and histone modification profiles of MCF7 cells are shown for the ±3 kb windows around the center of TCF7L2 peaks identified as proximal to RefSeq genes (upper graph) or distal to RefSeq genes (lower graph).

    Journal: Genome Biology

    Article Title: Cell type-specific binding patterns reveal that TCF7L2 can be tethered to the genome by association with GATA3

    doi: 10.1186/gb-2012-13-9-r52

    Figure Lengend Snippet: TCF7L2 binding sites are distal and enriched for active enhancer histone marks . (a) Shown for the TCF7L2 binding sites in the six cell types and for the 1,864 peaks commonly bound in all six cells is the percentage of TCF7L2 binding sites in different genomic regions (hg19) relative to the nearest transcription start site (TSS). (b) The percentage of active enhancer regions containing a TCF7L2 binding site; active enhancers were defined by taking the regions that have an overlap of H3K4me1 and H3K27ac ChIP-seq peaks for the given cell line. (c) Heatmaps of the ChIP-Seq tags for H3K27ac and H3K4me1 at TCF7L2-bound regions (±3 kb windows around the center of all TCF7L2 peaks) for each cell line were generated by k-means cluster analysis. (d) The average RNA polymerase II and histone modification profiles of MCF7 cells are shown for the ±3 kb windows around the center of TCF7L2 peaks identified as proximal to RefSeq genes (upper graph) or distal to RefSeq genes (lower graph).

    Article Snippet: To determine if we had identified the majority of the TCF7L2 peaks in each cell type, we performed a saturation analysis.

    Techniques: Binding Assay, Chromatin Immunoprecipitation, Generated, Modification

    Association of other motifs with TCF7L2 binding sites . (a,b) TCF7L2 binding sites unique to HepG2 cells (a) or MCF7 cells (b) were analyzed for the indicated motifs; the position of each motif is plotted relative to the center of the TCF7L2 binding site.

    Journal: Genome Biology

    Article Title: Cell type-specific binding patterns reveal that TCF7L2 can be tethered to the genome by association with GATA3

    doi: 10.1186/gb-2012-13-9-r52

    Figure Lengend Snippet: Association of other motifs with TCF7L2 binding sites . (a,b) TCF7L2 binding sites unique to HepG2 cells (a) or MCF7 cells (b) were analyzed for the indicated motifs; the position of each motif is plotted relative to the center of the TCF7L2 binding site.

    Article Snippet: To determine if we had identified the majority of the TCF7L2 peaks in each cell type, we performed a saturation analysis.

    Techniques: Binding Assay

    Association of TCF7L2 and GATA3 in MCF7 cells . (a) GATA3 ChIP-seq in MCF7 cells was performed, and peaks were called and then overlapped with the MCF7 cell type-specific TCF7L2 peaks; a comparison of TCF7L2 and GATA3 binding patterns near the CDT1 locus is shown. The hg19 genomic coordinates are chr16:88,861,964-88,880,233. (b) Peaks bound only by GATA3, only by TCF7L2, or by both factors were analyzed for the presence of GATA3 and TCF7L2 motifs. The GATA3 motif is found in sites bound by GATA3 only and in sites bound by both factors, whereas the TCF7L2 motif is found only in the sites bound only by TCF7L2 and not in the sites bound by both factors. (c) Depletion of GATA3 results in loss of TCF7L2 occupancy at sites bound by TCF7L2 and GATA3 sites but not at sites only bound by TCF7L2. MCF7 cells were transfected with siRNAs specific for TCF7L2 or GATA3 or control siRNAs. ChIP-qPCR assays were performed using antibodies specific for TCF7L2 (left panel) or GATA3 (right panel) using primers specific for peaks bound only by GATA3, only by TCF7L2, or by both factors. Shown are ChIP-qPCR results performed in triplicate and plotted with the standard error of two independent experiments. (d) Co-immunoprecipitation of endogenous GATA3 and FLAG-tagged TCF7L2 constructs from MCF7 cells. The left panel analyzes whole-cell extracts (WCE) and FLAG immunoprecipitation (FLAG IP) eluates from MCF7 cells transfected with the indicated FLAG-tagged plasmids; the membrane was incubated with both anti-FLAG and anti-GATA3 antibodies. Note that the GATA3 signal in input WCE extracts is quite weak and can generally only be visualized after concentration by immunoprecipitation. The right panel is a separate blot prepared in the same way (using the GATA antibody for immunoprecipitation), but does not include the WCE extracts. V, vector control; E, full length TCF7L2; EΔ, TCF7L2 lacking the amino terminus; B, TCF7L2 isoform lacking the carboxyl terminus; BΔ, TCF7L2 isoform lacking the amino and carboxyl termini.

    Journal: Genome Biology

    Article Title: Cell type-specific binding patterns reveal that TCF7L2 can be tethered to the genome by association with GATA3

    doi: 10.1186/gb-2012-13-9-r52

    Figure Lengend Snippet: Association of TCF7L2 and GATA3 in MCF7 cells . (a) GATA3 ChIP-seq in MCF7 cells was performed, and peaks were called and then overlapped with the MCF7 cell type-specific TCF7L2 peaks; a comparison of TCF7L2 and GATA3 binding patterns near the CDT1 locus is shown. The hg19 genomic coordinates are chr16:88,861,964-88,880,233. (b) Peaks bound only by GATA3, only by TCF7L2, or by both factors were analyzed for the presence of GATA3 and TCF7L2 motifs. The GATA3 motif is found in sites bound by GATA3 only and in sites bound by both factors, whereas the TCF7L2 motif is found only in the sites bound only by TCF7L2 and not in the sites bound by both factors. (c) Depletion of GATA3 results in loss of TCF7L2 occupancy at sites bound by TCF7L2 and GATA3 sites but not at sites only bound by TCF7L2. MCF7 cells were transfected with siRNAs specific for TCF7L2 or GATA3 or control siRNAs. ChIP-qPCR assays were performed using antibodies specific for TCF7L2 (left panel) or GATA3 (right panel) using primers specific for peaks bound only by GATA3, only by TCF7L2, or by both factors. Shown are ChIP-qPCR results performed in triplicate and plotted with the standard error of two independent experiments. (d) Co-immunoprecipitation of endogenous GATA3 and FLAG-tagged TCF7L2 constructs from MCF7 cells. The left panel analyzes whole-cell extracts (WCE) and FLAG immunoprecipitation (FLAG IP) eluates from MCF7 cells transfected with the indicated FLAG-tagged plasmids; the membrane was incubated with both anti-FLAG and anti-GATA3 antibodies. Note that the GATA3 signal in input WCE extracts is quite weak and can generally only be visualized after concentration by immunoprecipitation. The right panel is a separate blot prepared in the same way (using the GATA antibody for immunoprecipitation), but does not include the WCE extracts. V, vector control; E, full length TCF7L2; EΔ, TCF7L2 lacking the amino terminus; B, TCF7L2 isoform lacking the carboxyl terminus; BΔ, TCF7L2 isoform lacking the amino and carboxyl termini.

    Article Snippet: To determine if we had identified the majority of the TCF7L2 peaks in each cell type, we performed a saturation analysis.

    Techniques: Chromatin Immunoprecipitation, Binding Assay, Transfection, Real-time Polymerase Chain Reaction, Immunoprecipitation, Construct, Incubation, Concentration Assay, Plasmid Preparation

    ChIP-seq analysis of TCF7L2 in six different human cell lines . Shown is the distribution of TCF7L2 binding within ±3 kb windows around distinct genomic regions (n = 3,000) bound by TCF7L2 in a given cell type. ChIP-seq tags for each cell line were each aligned with respect to the center of the combined top 500 peaks from each dataset and clustered by genomic position.

    Journal: Genome Biology

    Article Title: Cell type-specific binding patterns reveal that TCF7L2 can be tethered to the genome by association with GATA3

    doi: 10.1186/gb-2012-13-9-r52

    Figure Lengend Snippet: ChIP-seq analysis of TCF7L2 in six different human cell lines . Shown is the distribution of TCF7L2 binding within ±3 kb windows around distinct genomic regions (n = 3,000) bound by TCF7L2 in a given cell type. ChIP-seq tags for each cell line were each aligned with respect to the center of the combined top 500 peaks from each dataset and clustered by genomic position.

    Article Snippet: To determine if we had identified the majority of the TCF7L2 peaks in each cell type, we performed a saturation analysis.

    Techniques: Chromatin Immunoprecipitation, Binding Assay

    Expression of DN-TCF7L2 reduced SMK-17-induced apoptosis in HCT 116 cells. HCT 116 cells were transfected with DN-TCF7L2. (A) Luciferase activity was monitored through co-transfection with the TOPFlash and Renilla luciferase plasmids. (B) HCT 116 cells expressing DN-TCF7L2 were treated with the indicated concentrations of SMK-17 for 48 h and stained with annexin V/APC and 7-AAD. Cells were analyzed by flow cytometry with gates including only EGFP-positive cells.

    Journal: Scientific Reports

    Article Title: SMK-17, a MEK1/2-specific inhibitor, selectively induces apoptosis in β-catenin-mutated tumors

    doi: 10.1038/srep08155

    Figure Lengend Snippet: Expression of DN-TCF7L2 reduced SMK-17-induced apoptosis in HCT 116 cells. HCT 116 cells were transfected with DN-TCF7L2. (A) Luciferase activity was monitored through co-transfection with the TOPFlash and Renilla luciferase plasmids. (B) HCT 116 cells expressing DN-TCF7L2 were treated with the indicated concentrations of SMK-17 for 48 h and stained with annexin V/APC and 7-AAD. Cells were analyzed by flow cytometry with gates including only EGFP-positive cells.

    Article Snippet: In the vector-transfection experiments, we co-transfected the EGFP expression vector as a transfectant marker with DN-TCF7L2 (Merck Millipore, Billerica, MA) or an active, mutated human β-catenin (S37A and S45A) expression vector (Daiichi Sankyo Co. Ltd.).

    Techniques: Expressing, Transfection, Luciferase, Activity Assay, Cotransfection, Staining, Flow Cytometry, Cytometry

    Activation of Wnt/β-catenin signaling by Wnt3a induced apoptosis in A375 cells. (A) A375 cells were co-transfected with the TOPFlash and Renilla luciferase plasmids. Transfected cells were treated with 50 ng/mL Wnt3a for 24 h, and their luciferase activity was measured. Wnt signaling activity in terms of TCF7L2 transcription was monitored by using the TOPFlash/continuous Renilla luciferase assay. The ratio of the firefly luciferase intensity of TOPFlash to that of Renilla luciferase was used as an indicator of TCF7L2 transcriptional activity. (B) Cells were treated with either DMSO or with 10 μM SMK-17, with or without 50 ng/mL recombinant Wnt3a, for 48 h. Cell lysates were probed for ERK1/2, phospho-ERK1/2 (p-ERK1/2), and cleaved PARP via western blot. Cleaved PARP was observed after combination treatment with SMK-17 and Wnt3a.

    Journal: Scientific Reports

    Article Title: SMK-17, a MEK1/2-specific inhibitor, selectively induces apoptosis in β-catenin-mutated tumors

    doi: 10.1038/srep08155

    Figure Lengend Snippet: Activation of Wnt/β-catenin signaling by Wnt3a induced apoptosis in A375 cells. (A) A375 cells were co-transfected with the TOPFlash and Renilla luciferase plasmids. Transfected cells were treated with 50 ng/mL Wnt3a for 24 h, and their luciferase activity was measured. Wnt signaling activity in terms of TCF7L2 transcription was monitored by using the TOPFlash/continuous Renilla luciferase assay. The ratio of the firefly luciferase intensity of TOPFlash to that of Renilla luciferase was used as an indicator of TCF7L2 transcriptional activity. (B) Cells were treated with either DMSO or with 10 μM SMK-17, with or without 50 ng/mL recombinant Wnt3a, for 48 h. Cell lysates were probed for ERK1/2, phospho-ERK1/2 (p-ERK1/2), and cleaved PARP via western blot. Cleaved PARP was observed after combination treatment with SMK-17 and Wnt3a.

    Article Snippet: In the vector-transfection experiments, we co-transfected the EGFP expression vector as a transfectant marker with DN-TCF7L2 (Merck Millipore, Billerica, MA) or an active, mutated human β-catenin (S37A and S45A) expression vector (Daiichi Sankyo Co. Ltd.).

    Techniques: Activation Assay, Transfection, Luciferase, Activity Assay, Recombinant, Western Blot

    A-C. Stained cells within the arcuate hypothalamic nucleus. A. NPY-stained cells showing immunoreactive fibers extending beyond the nucleus into the region adjacent to the arcuate nucleus. B. TCF7L2 stained nuclei of arcuate neurons. C. Agouti-related

    Journal: Journal of chemical neuroanatomy

    Article Title: Review of the Neuroanatomic Landscape Implicated in Glucose Sensing and Regulation of Nutrient Signaling: Immunophenotypic Localization of Diabetes Gene Tcf7l2 in the Developing Murine Brain

    doi: 10.1016/j.jchemneu.2012.06.002

    Figure Lengend Snippet: A-C. Stained cells within the arcuate hypothalamic nucleus. A. NPY-stained cells showing immunoreactive fibers extending beyond the nucleus into the region adjacent to the arcuate nucleus. B. TCF7L2 stained nuclei of arcuate neurons. C. Agouti-related

    Article Snippet: As shown in taken from a series of coronal brain sections, the immunohistochemical localization of TCF7L2 is relatively widespread and varies in its expression within neuronal populations and regions of the brain in the wild type and het progeny at E18.5.

    Techniques: Staining

    A-C. Labeled cells of the lateral preoptic nucleus(A), medial preoptic nucleus(B) and septal neuroepithelium(C). 20× D. Parasagittal view of a P1 Tcf7l2 +/- head showing the distribution of TCF7L2 within the thalamus and midbrain regions. 1×

    Journal: Journal of chemical neuroanatomy

    Article Title: Review of the Neuroanatomic Landscape Implicated in Glucose Sensing and Regulation of Nutrient Signaling: Immunophenotypic Localization of Diabetes Gene Tcf7l2 in the Developing Murine Brain

    doi: 10.1016/j.jchemneu.2012.06.002

    Figure Lengend Snippet: A-C. Labeled cells of the lateral preoptic nucleus(A), medial preoptic nucleus(B) and septal neuroepithelium(C). 20× D. Parasagittal view of a P1 Tcf7l2 +/- head showing the distribution of TCF7L2 within the thalamus and midbrain regions. 1×

    Article Snippet: As shown in taken from a series of coronal brain sections, the immunohistochemical localization of TCF7L2 is relatively widespread and varies in its expression within neuronal populations and regions of the brain in the wild type and het progeny at E18.5.

    Techniques: Labeling

    A-I. Medullary to midbrain coronal sections of an E18.5 Tcf7l2 +/+ mouse showing the widespread distribution of TCF7L2 peptide. A-F show the distribution of the peptide within nuclei of the hindbrain while G-I are midbrain sections illustrating the prevalence

    Journal: Journal of chemical neuroanatomy

    Article Title: Review of the Neuroanatomic Landscape Implicated in Glucose Sensing and Regulation of Nutrient Signaling: Immunophenotypic Localization of Diabetes Gene Tcf7l2 in the Developing Murine Brain

    doi: 10.1016/j.jchemneu.2012.06.002

    Figure Lengend Snippet: A-I. Medullary to midbrain coronal sections of an E18.5 Tcf7l2 +/+ mouse showing the widespread distribution of TCF7L2 peptide. A-F show the distribution of the peptide within nuclei of the hindbrain while G-I are midbrain sections illustrating the prevalence

    Article Snippet: As shown in taken from a series of coronal brain sections, the immunohistochemical localization of TCF7L2 is relatively widespread and varies in its expression within neuronal populations and regions of the brain in the wild type and het progeny at E18.5.

    Techniques:

    Cumulative weight gain ( A ), absolute body weight ( B ), final body weight ( C ), and cumulative food intake ( D ) during and after 12 weeks of the HFD ( n = 14 ΔE11-TCF7L2, n = 16 LoxP controls). * P

    Journal: Diabetes

    Article Title: The Diabetes Gene and Wnt Pathway Effector TCF7L2 Regulates Adipocyte Development and Function

    doi: 10.2337/db17-0318

    Figure Lengend Snippet: Cumulative weight gain ( A ), absolute body weight ( B ), final body weight ( C ), and cumulative food intake ( D ) during and after 12 weeks of the HFD ( n = 14 ΔE11-TCF7L2, n = 16 LoxP controls). * P

    Article Snippet: Total TCF7L2 mRNA, detected using a TaqMan probe designed around exons 10 and 11 (assay ID: Hs01009038_m1) was significantly decreased in IGT subjects ( ) as was the expression of a “full adipose tissue variant” ( ) of TCF7L2 that incorporates exons 12 and 13 ( ).

    Techniques:

    A : Schematic shows the targeting strategy for the mouse Tcf7l2 ]) B : LoxP sites were placed around exon 11, which results in the deletion of exon 11, and not exon 6, containing

    Journal: Diabetes

    Article Title: The Diabetes Gene and Wnt Pathway Effector TCF7L2 Regulates Adipocyte Development and Function

    doi: 10.2337/db17-0318

    Figure Lengend Snippet: A : Schematic shows the targeting strategy for the mouse Tcf7l2 ]) B : LoxP sites were placed around exon 11, which results in the deletion of exon 11, and not exon 6, containing

    Article Snippet: Total TCF7L2 mRNA, detected using a TaqMan probe designed around exons 10 and 11 (assay ID: Hs01009038_m1) was significantly decreased in IGT subjects ( ) as was the expression of a “full adipose tissue variant” ( ) of TCF7L2 that incorporates exons 12 and 13 ( ).

    Techniques:

    A : iWAT in male ΔE11-TCF7L2 mice at 3 and 6 months of age ( n = 8–10 ΔE11-TCF7L2, n = 8–10 control mice) and iWAT and pgWAT after HFD ( n = 12 ΔE11-TCF7L2, n = 14 control mice). ** P

    Journal: Diabetes

    Article Title: The Diabetes Gene and Wnt Pathway Effector TCF7L2 Regulates Adipocyte Development and Function

    doi: 10.2337/db17-0318

    Figure Lengend Snippet: A : iWAT in male ΔE11-TCF7L2 mice at 3 and 6 months of age ( n = 8–10 ΔE11-TCF7L2, n = 8–10 control mice) and iWAT and pgWAT after HFD ( n = 12 ΔE11-TCF7L2, n = 14 control mice). ** P

    Article Snippet: Total TCF7L2 mRNA, detected using a TaqMan probe designed around exons 10 and 11 (assay ID: Hs01009038_m1) was significantly decreased in IGT subjects ( ) as was the expression of a “full adipose tissue variant” ( ) of TCF7L2 that incorporates exons 12 and 13 ( ).

    Techniques: Mouse Assay

    A : Representative images of differentiating 3T3-L1 adipocytes at the time of induction, day 0 (D0), and at D4 and D10 after initiation of adipogenesis (scale bar = 100 µm). B and C : A representative Western blot of TCF7L2 during adipogenesis,

    Journal: Diabetes

    Article Title: The Diabetes Gene and Wnt Pathway Effector TCF7L2 Regulates Adipocyte Development and Function

    doi: 10.2337/db17-0318

    Figure Lengend Snippet: A : Representative images of differentiating 3T3-L1 adipocytes at the time of induction, day 0 (D0), and at D4 and D10 after initiation of adipogenesis (scale bar = 100 µm). B and C : A representative Western blot of TCF7L2 during adipogenesis,

    Article Snippet: Total TCF7L2 mRNA, detected using a TaqMan probe designed around exons 10 and 11 (assay ID: Hs01009038_m1) was significantly decreased in IGT subjects ( ) as was the expression of a “full adipose tissue variant” ( ) of TCF7L2 that incorporates exons 12 and 13 ( ).

    Techniques: Western Blot

    Characterization of TCF7L2 up-regulation in response to 1,25(OH) 2 D 3 . ( A ) qPCR analysis of CaCo2 cells transfected and treated as described in Figure 2D and 2E . TCF7L2 (left panel) and CYP24A1 (right panel) transcripts. Data are normalized to GAPDH and plotted relative to each EtOH-treated control. p-values are derived from student's t-test comparison between EtOH and D 3 controls: * p

    Journal: PLoS ONE

    Article Title: Control of TCF-4 Expression by VDR and Vitamin D in the Mouse Mammary Gland and Colorectal Cancer Cell Lines

    doi: 10.1371/journal.pone.0007872

    Figure Lengend Snippet: Characterization of TCF7L2 up-regulation in response to 1,25(OH) 2 D 3 . ( A ) qPCR analysis of CaCo2 cells transfected and treated as described in Figure 2D and 2E . TCF7L2 (left panel) and CYP24A1 (right panel) transcripts. Data are normalized to GAPDH and plotted relative to each EtOH-treated control. p-values are derived from student's t-test comparison between EtOH and D 3 controls: * p

    Article Snippet: Subsequent cDNA was used in a quantitative amplification reaction using Applied Biosystems TaqMan Universal Master Mix (Cat #4304437) with the following primer/probes: human CYP24A1 (Hs00167999_m1), human TCF7L2 (Hs01009053_m1), human GAPDH (Hs99999905_m1), mouse TCF7L2 (Mm01261075_m1), mouse β-actin (Mm01205647_g1).

    Techniques: Real-time Polymerase Chain Reaction, Transfection, Derivative Assay

    Mouse TCF7L2 promoter reporter constructs respond to VDR or 1,25(OH) 2 D 3 . ( A ) Two mouse TCF7L2 promoter constructs were cloned upstream of a luciferase reporter. The four putative VDRE locations relative to the translation start site are denoted, and all VDREs are encoded on the non-coding strand. The −177 and −187 VDREs are overlapping and share a half-site between them. The −1037 construct (-1037-luc) contains the region from −1 to −1037 relative to the translation start site and possesses the −177/−187 VDRE, only, while the −2068 reporter (-2068-luc) contains the region between −96 and −2068 relative to the translation start site of the mouse TCF7L2 promoter, and possesses all four putative VDREs. ( B ) CaCo2 cells were transfected for 24 hours with Renilla and reporters as indicated as well as different amounts of p53. Data are normalized to Renilla and to empty reporter expression. RLU = Relative Light Units. ( C ) -1038-Luc or empty vector constructs were transfected into CaCo2 cells with Renilla and different amounts of VDR, as indicated. 24 hours after transfection, cells were treated for an additional 24 hours with 10 −7 M 1,25(OH) 2 D 3 or EtOH vehicle control. Error bars represent SEM. Statistics were derived from two-way ANOVA for differences between EtOH- and 1,25(OH) 2 D 3 -treated samples: * p

    Journal: PLoS ONE

    Article Title: Control of TCF-4 Expression by VDR and Vitamin D in the Mouse Mammary Gland and Colorectal Cancer Cell Lines

    doi: 10.1371/journal.pone.0007872

    Figure Lengend Snippet: Mouse TCF7L2 promoter reporter constructs respond to VDR or 1,25(OH) 2 D 3 . ( A ) Two mouse TCF7L2 promoter constructs were cloned upstream of a luciferase reporter. The four putative VDRE locations relative to the translation start site are denoted, and all VDREs are encoded on the non-coding strand. The −177 and −187 VDREs are overlapping and share a half-site between them. The −1037 construct (-1037-luc) contains the region from −1 to −1037 relative to the translation start site and possesses the −177/−187 VDRE, only, while the −2068 reporter (-2068-luc) contains the region between −96 and −2068 relative to the translation start site of the mouse TCF7L2 promoter, and possesses all four putative VDREs. ( B ) CaCo2 cells were transfected for 24 hours with Renilla and reporters as indicated as well as different amounts of p53. Data are normalized to Renilla and to empty reporter expression. RLU = Relative Light Units. ( C ) -1038-Luc or empty vector constructs were transfected into CaCo2 cells with Renilla and different amounts of VDR, as indicated. 24 hours after transfection, cells were treated for an additional 24 hours with 10 −7 M 1,25(OH) 2 D 3 or EtOH vehicle control. Error bars represent SEM. Statistics were derived from two-way ANOVA for differences between EtOH- and 1,25(OH) 2 D 3 -treated samples: * p

    Article Snippet: Subsequent cDNA was used in a quantitative amplification reaction using Applied Biosystems TaqMan Universal Master Mix (Cat #4304437) with the following primer/probes: human CYP24A1 (Hs00167999_m1), human TCF7L2 (Hs01009053_m1), human GAPDH (Hs99999905_m1), mouse TCF7L2 (Mm01261075_m1), mouse β-actin (Mm01205647_g1).

    Techniques: Construct, Clone Assay, Luciferase, Transfection, Expressing, Plasmid Preparation, Derivative Assay

    Cycloheximide treatment abolishes 1,25(OH) 2 D 3 -induced TCF7L2 mRNA induction. CaCo2 cells were pre-treated for 30 minutes with different concentrations of the protein synthesis inhibitor Cycloheximide (CHX) before addition of 10 −7 M 1,25(OH) 2 D 3 or EtOH for 24 hours, as indicated. Analysis of mRNA abundance of TCF7L2 (top panel) and DKK4 (bottom panel) was assayed by qPCR. Error bars represent SEM. * p

    Journal: PLoS ONE

    Article Title: Control of TCF-4 Expression by VDR and Vitamin D in the Mouse Mammary Gland and Colorectal Cancer Cell Lines

    doi: 10.1371/journal.pone.0007872

    Figure Lengend Snippet: Cycloheximide treatment abolishes 1,25(OH) 2 D 3 -induced TCF7L2 mRNA induction. CaCo2 cells were pre-treated for 30 minutes with different concentrations of the protein synthesis inhibitor Cycloheximide (CHX) before addition of 10 −7 M 1,25(OH) 2 D 3 or EtOH for 24 hours, as indicated. Analysis of mRNA abundance of TCF7L2 (top panel) and DKK4 (bottom panel) was assayed by qPCR. Error bars represent SEM. * p

    Article Snippet: Subsequent cDNA was used in a quantitative amplification reaction using Applied Biosystems TaqMan Universal Master Mix (Cat #4304437) with the following primer/probes: human CYP24A1 (Hs00167999_m1), human TCF7L2 (Hs01009053_m1), human GAPDH (Hs99999905_m1), mouse TCF7L2 (Mm01261075_m1), mouse β-actin (Mm01205647_g1).

    Techniques: Real-time Polymerase Chain Reaction

    VDR145 +/+ cells have more basal TCF-4 than VDRK240 −/− cells. ( A ) Western blot of whole cell lysates, in duplicate, from VDR145 +/+ (VDR +/+) cells (lanes 1 and 2) and VDRK240 −/− (VDR −/−) cells (lanes 3 and 4) probed for TCF-4, VDR and GAPDH. ( B ) Pull-down of proteins in VDR +/+ and VDR −/− lysates with GST-tagged β-catenin constructs and probed for TCF-4. GST-WT-β-catenin (GST-β-Cat) is used in lanes 1 and 2. Mutated β-catenin that will not effectively bind TCF/LEF proteins (GST-dTCF-β-Cat) is used in lanes 3 and 4. Beads alone are used in lanes 5 and 6. ( C ) OT activity in VDR +/+ and VDR −/− cells transfected with Renilla and VP16-β-catenin. Data are normalized to Renilla . Error bars represent SEM. p-values represent student's t-test for significan differences between cell lines. RLU: Relative Light Units ( D ) OT activity in VDR +/+ and VDR −/− cells transfected with Renilla , VP16-β-catenin and with or without a TCF7L2 plasmid. Error bars represent SEM. Statistics are student's t-test for differences between transfected and control-transfected cells. *p

    Journal: PLoS ONE

    Article Title: Control of TCF-4 Expression by VDR and Vitamin D in the Mouse Mammary Gland and Colorectal Cancer Cell Lines

    doi: 10.1371/journal.pone.0007872

    Figure Lengend Snippet: VDR145 +/+ cells have more basal TCF-4 than VDRK240 −/− cells. ( A ) Western blot of whole cell lysates, in duplicate, from VDR145 +/+ (VDR +/+) cells (lanes 1 and 2) and VDRK240 −/− (VDR −/−) cells (lanes 3 and 4) probed for TCF-4, VDR and GAPDH. ( B ) Pull-down of proteins in VDR +/+ and VDR −/− lysates with GST-tagged β-catenin constructs and probed for TCF-4. GST-WT-β-catenin (GST-β-Cat) is used in lanes 1 and 2. Mutated β-catenin that will not effectively bind TCF/LEF proteins (GST-dTCF-β-Cat) is used in lanes 3 and 4. Beads alone are used in lanes 5 and 6. ( C ) OT activity in VDR +/+ and VDR −/− cells transfected with Renilla and VP16-β-catenin. Data are normalized to Renilla . Error bars represent SEM. p-values represent student's t-test for significan differences between cell lines. RLU: Relative Light Units ( D ) OT activity in VDR +/+ and VDR −/− cells transfected with Renilla , VP16-β-catenin and with or without a TCF7L2 plasmid. Error bars represent SEM. Statistics are student's t-test for differences between transfected and control-transfected cells. *p

    Article Snippet: Subsequent cDNA was used in a quantitative amplification reaction using Applied Biosystems TaqMan Universal Master Mix (Cat #4304437) with the following primer/probes: human CYP24A1 (Hs00167999_m1), human TCF7L2 (Hs01009053_m1), human GAPDH (Hs99999905_m1), mouse TCF7L2 (Mm01261075_m1), mouse β-actin (Mm01205647_g1).

    Techniques: Western Blot, Construct, Activity Assay, Transfection, Plasmid Preparation

    Putative VDREs within the TCF7L2 Promoter are not Responsible for 1,25(OH) 2 D 3 -VDR-mediated increase of TCF7L2. ( A ) Graphical representation of the four putative VDREs within the first ∼1500 base-pairs of the mouse TCF7L2 promoter and their sequences. DR3: direct repeat interspersed by three nucleotides. 2xDR4: two, overlapping direct repeats interspersed by four nucleotides. Although the putative VDREs are encoded on the non-coding strand, their sequences are written in reverse-complement such that the VDREs can be identified as conforming to the RGKTSA consensus. ( B ) -2068-luc construct containing all three sets of half-site mutations (d1502/d1153/d177) was transfected into CaCo2 cells and treated with ligand as described in Figure 4C with only 3 concentrations of VDR (low, medium and high). Error bars represent SEM. Statistics represent analysis using two-way ANOVA: *p

    Journal: PLoS ONE

    Article Title: Control of TCF-4 Expression by VDR and Vitamin D in the Mouse Mammary Gland and Colorectal Cancer Cell Lines

    doi: 10.1371/journal.pone.0007872

    Figure Lengend Snippet: Putative VDREs within the TCF7L2 Promoter are not Responsible for 1,25(OH) 2 D 3 -VDR-mediated increase of TCF7L2. ( A ) Graphical representation of the four putative VDREs within the first ∼1500 base-pairs of the mouse TCF7L2 promoter and their sequences. DR3: direct repeat interspersed by three nucleotides. 2xDR4: two, overlapping direct repeats interspersed by four nucleotides. Although the putative VDREs are encoded on the non-coding strand, their sequences are written in reverse-complement such that the VDREs can be identified as conforming to the RGKTSA consensus. ( B ) -2068-luc construct containing all three sets of half-site mutations (d1502/d1153/d177) was transfected into CaCo2 cells and treated with ligand as described in Figure 4C with only 3 concentrations of VDR (low, medium and high). Error bars represent SEM. Statistics represent analysis using two-way ANOVA: *p

    Article Snippet: Subsequent cDNA was used in a quantitative amplification reaction using Applied Biosystems TaqMan Universal Master Mix (Cat #4304437) with the following primer/probes: human CYP24A1 (Hs00167999_m1), human TCF7L2 (Hs01009053_m1), human GAPDH (Hs99999905_m1), mouse TCF7L2 (Mm01261075_m1), mouse β-actin (Mm01205647_g1).

    Techniques: Construct, Transfection

    MUC1-C cytoplasmic domain associates with TCF7L2. A , lysates from ZR-75-1 ( left ) and MCF-7 ( right ) breast cancer cells were precipitated with anti-TCF7L2 or, as a control, non-immune IgG. The precipitates were immunoblotted with the indicated antibodies.

    Journal: The Journal of Biological Chemistry

    Article Title: MUC1-C Oncoprotein Induces TCF7L2 Transcription Factor Activation and Promotes Cyclin D1 Expression in Human Breast Cancer Cells

    doi: 10.1074/jbc.M111.323311

    Figure Lengend Snippet: MUC1-C cytoplasmic domain associates with TCF7L2. A , lysates from ZR-75-1 ( left ) and MCF-7 ( right ) breast cancer cells were precipitated with anti-TCF7L2 or, as a control, non-immune IgG. The precipitates were immunoblotted with the indicated antibodies.

    Article Snippet: Soluble proteins (500 μg) were precipitated with anti-TCF7L2 (sc-13027, 1:50; Santa Cruz Biotechnology).

    Techniques:

    MUC1-C occupies the cyclin D1 promoter in a complex with TCF7L2. A , schematic representation of the cyclin D1 promoter with positioning of the TCF binding sites. B and C , soluble chromatin from ZR-75-1 ( B ) and MCF-7 ( C ) cells was precipitated with anti-TCF7L2

    Journal: The Journal of Biological Chemistry

    Article Title: MUC1-C Oncoprotein Induces TCF7L2 Transcription Factor Activation and Promotes Cyclin D1 Expression in Human Breast Cancer Cells

    doi: 10.1074/jbc.M111.323311

    Figure Lengend Snippet: MUC1-C occupies the cyclin D1 promoter in a complex with TCF7L2. A , schematic representation of the cyclin D1 promoter with positioning of the TCF binding sites. B and C , soluble chromatin from ZR-75-1 ( B ) and MCF-7 ( C ) cells was precipitated with anti-TCF7L2

    Article Snippet: Soluble proteins (500 μg) were precipitated with anti-TCF7L2 (sc-13027, 1:50; Santa Cruz Biotechnology).

    Techniques: Binding Assay

    MUC1-CD CQC motif confers binding to TCF7L2. A , GST, GST-MUC1-CD(1–45), or GST-MUC1-CD(46–72) were incubated with recombinant TCF7L2. Adsorbates were immunoblotted ( IB ) with anti-TCF7L2. Input of the GST proteins was assessed by Coomassie

    Journal: The Journal of Biological Chemistry

    Article Title: MUC1-C Oncoprotein Induces TCF7L2 Transcription Factor Activation and Promotes Cyclin D1 Expression in Human Breast Cancer Cells

    doi: 10.1074/jbc.M111.323311

    Figure Lengend Snippet: MUC1-CD CQC motif confers binding to TCF7L2. A , GST, GST-MUC1-CD(1–45), or GST-MUC1-CD(46–72) were incubated with recombinant TCF7L2. Adsorbates were immunoblotted ( IB ) with anti-TCF7L2. Input of the GST proteins was assessed by Coomassie

    Article Snippet: Soluble proteins (500 μg) were precipitated with anti-TCF7L2 (sc-13027, 1:50; Santa Cruz Biotechnology).

    Techniques: Binding Assay, Incubation, Recombinant

    Inhibition of MUC1-C decreases cyclin D1 expression. A , amino acid sequences of the GO-201 and control CP-1 peptides. GST-TCF7L2 was incubated with MUC1-CD in the absence ( Control ) and presence of GO-201 or CP-1. Adsorbates were immunoblotted with anti-MUC1-C.

    Journal: The Journal of Biological Chemistry

    Article Title: MUC1-C Oncoprotein Induces TCF7L2 Transcription Factor Activation and Promotes Cyclin D1 Expression in Human Breast Cancer Cells

    doi: 10.1074/jbc.M111.323311

    Figure Lengend Snippet: Inhibition of MUC1-C decreases cyclin D1 expression. A , amino acid sequences of the GO-201 and control CP-1 peptides. GST-TCF7L2 was incubated with MUC1-CD in the absence ( Control ) and presence of GO-201 or CP-1. Adsorbates were immunoblotted with anti-MUC1-C.

    Article Snippet: Soluble proteins (500 μg) were precipitated with anti-TCF7L2 (sc-13027, 1:50; Santa Cruz Biotechnology).

    Techniques: Inhibition, Expressing, Incubation

    MUC1-C promotes TCF7L2 activation of the cyclin D1 promoter. A , the indicated ZR-75-1 ( left ) and MCF-7 ( right ) cells were transfected with pGL3 ( shaded bars ) or pGL3 expressing pcycD1(−166)-Luc ( solid bars ) for 48 h and then assayed for luciferase

    Journal: The Journal of Biological Chemistry

    Article Title: MUC1-C Oncoprotein Induces TCF7L2 Transcription Factor Activation and Promotes Cyclin D1 Expression in Human Breast Cancer Cells

    doi: 10.1074/jbc.M111.323311

    Figure Lengend Snippet: MUC1-C promotes TCF7L2 activation of the cyclin D1 promoter. A , the indicated ZR-75-1 ( left ) and MCF-7 ( right ) cells were transfected with pGL3 ( shaded bars ) or pGL3 expressing pcycD1(−166)-Luc ( solid bars ) for 48 h and then assayed for luciferase

    Article Snippet: Soluble proteins (500 μg) were precipitated with anti-TCF7L2 (sc-13027, 1:50; Santa Cruz Biotechnology).

    Techniques: Activation Assay, Transfection, Expressing, Luciferase

    MUC1-CD binds directly to the TCF7L2 CGPCRRK motif in the E-tail. A and B , GST-TCF7L2(295–476) ( A ) or GST-TCF7L2(477–596) ( B ) was incubated with MUC1-CD or MUC1-CD(AQA). The adsorbates were immunoblotted with anti-MUC1-C. Input of the

    Journal: The Journal of Biological Chemistry

    Article Title: MUC1-C Oncoprotein Induces TCF7L2 Transcription Factor Activation and Promotes Cyclin D1 Expression in Human Breast Cancer Cells

    doi: 10.1074/jbc.M111.323311

    Figure Lengend Snippet: MUC1-CD binds directly to the TCF7L2 CGPCRRK motif in the E-tail. A and B , GST-TCF7L2(295–476) ( A ) or GST-TCF7L2(477–596) ( B ) was incubated with MUC1-CD or MUC1-CD(AQA). The adsorbates were immunoblotted with anti-MUC1-C. Input of the

    Article Snippet: Soluble proteins (500 μg) were precipitated with anti-TCF7L2 (sc-13027, 1:50; Santa Cruz Biotechnology).

    Techniques: Incubation

    Cumulative weight gain ( A ), absolute body weight ( B ), final body weight ( C ), and cumulative food intake ( D ) during and after 12 weeks of the HFD ( n = 14 ΔE11-TCF7L2, n = 16 LoxP controls). * P

    Journal: Diabetes

    Article Title: The Diabetes Gene and Wnt Pathway Effector TCF7L2 Regulates Adipocyte Development and Function

    doi: 10.2337/db17-0318

    Figure Lengend Snippet: Cumulative weight gain ( A ), absolute body weight ( B ), final body weight ( C ), and cumulative food intake ( D ) during and after 12 weeks of the HFD ( n = 14 ΔE11-TCF7L2, n = 16 LoxP controls). * P

    Article Snippet: We measured mouse Tcf7l2 mRNA using a TaqMan probe that spans exon 5 and 6 of full length Tcf7l2 and detects all mRNA variants (assay ID: Mm00501505_m1).

    Techniques:

    A : Schematic shows the targeting strategy for the mouse Tcf7l2 locus used in this study. (Reprinted with permission from van Es et al. [ 19 ]) B : LoxP sites were placed around exon 11, which results in the deletion of exon 11, and not exon 6, containing mRNA transcripts, as determined by quantitative real-time PCR on isolated adipocytes ( n = 7). *** P

    Journal: Diabetes

    Article Title: The Diabetes Gene and Wnt Pathway Effector TCF7L2 Regulates Adipocyte Development and Function

    doi: 10.2337/db17-0318

    Figure Lengend Snippet: A : Schematic shows the targeting strategy for the mouse Tcf7l2 locus used in this study. (Reprinted with permission from van Es et al. [ 19 ]) B : LoxP sites were placed around exon 11, which results in the deletion of exon 11, and not exon 6, containing mRNA transcripts, as determined by quantitative real-time PCR on isolated adipocytes ( n = 7). *** P

    Article Snippet: We measured mouse Tcf7l2 mRNA using a TaqMan probe that spans exon 5 and 6 of full length Tcf7l2 and detects all mRNA variants (assay ID: Mm00501505_m1).

    Techniques: Real-time Polymerase Chain Reaction, Isolation

    A : iWAT in male ΔE11-TCF7L2 mice at 3 and 6 months of age ( n = 8–10 ΔE11-TCF7L2, n = 8–10 control mice) and iWAT and pgWAT after HFD ( n = 12 ΔE11-TCF7L2, n = 14 control mice). ** P

    Journal: Diabetes

    Article Title: The Diabetes Gene and Wnt Pathway Effector TCF7L2 Regulates Adipocyte Development and Function

    doi: 10.2337/db17-0318

    Figure Lengend Snippet: A : iWAT in male ΔE11-TCF7L2 mice at 3 and 6 months of age ( n = 8–10 ΔE11-TCF7L2, n = 8–10 control mice) and iWAT and pgWAT after HFD ( n = 12 ΔE11-TCF7L2, n = 14 control mice). ** P

    Article Snippet: We measured mouse Tcf7l2 mRNA using a TaqMan probe that spans exon 5 and 6 of full length Tcf7l2 and detects all mRNA variants (assay ID: Mm00501505_m1).

    Techniques: Mouse Assay

    A : Representative images of differentiating 3T3-L1 adipocytes at the time of induction, day 0 (D0), and at D4 and D10 after initiation of adipogenesis (scale bar = 100 µm). B and C : A representative Western blot of TCF7L2 during adipogenesis, examined using an antibody that recognizes an epitope around Leu331 and all variants of TCF7L2. The expression of the short (58 kDa) TCF7L2 protein isoform was higher than the long (78 kDa) isoform during adipogenesis. Total protein load, assessed by Ponceau-S staining, was used for normalization ( n = 5 independent experiments). *** P

    Journal: Diabetes

    Article Title: The Diabetes Gene and Wnt Pathway Effector TCF7L2 Regulates Adipocyte Development and Function

    doi: 10.2337/db17-0318

    Figure Lengend Snippet: A : Representative images of differentiating 3T3-L1 adipocytes at the time of induction, day 0 (D0), and at D4 and D10 after initiation of adipogenesis (scale bar = 100 µm). B and C : A representative Western blot of TCF7L2 during adipogenesis, examined using an antibody that recognizes an epitope around Leu331 and all variants of TCF7L2. The expression of the short (58 kDa) TCF7L2 protein isoform was higher than the long (78 kDa) isoform during adipogenesis. Total protein load, assessed by Ponceau-S staining, was used for normalization ( n = 5 independent experiments). *** P

    Article Snippet: We measured mouse Tcf7l2 mRNA using a TaqMan probe that spans exon 5 and 6 of full length Tcf7l2 and detects all mRNA variants (assay ID: Mm00501505_m1).

    Techniques: Western Blot, Expressing, Staining

    The TCF/LEF family of transcription factors bind the insertion allele of rs386772267 in an allele-specific manner. (A) Electromobility shift assay (EMSA) of rs386772267 deletion and insertion allele probes incubated with PANC-1 nuclear extract with and without unlabelled allelic competitors reveals an allele-specific band for the insertion probe (indicated by the arrow). (B) Alignment of binding sequence motifs for TCF/LEF transcription factors against the insertion probe (insertion allele designated in red). Similarity scores indicate similarity of aligned insertion allele probe to binding sequence matrices for the respective TFs as calculated by Genomatix MatInspector TM . (C and D) The ratios for medium and light labelled proteins bound to ( C ) insertion allele vs. scrambled insertion or ( D ) vs. deletion allele probes. Isotopic labeling for the y-axis experiments were reversed compared to those of the x-axis. Proteins indicated in red (TCF7L2 and SNRPA1) showed consistent preferential binding to the insertion allele vs. both scrambled insertion and deletion allele probes. (E) The insertion probe’s allele-specific EMSA band (indicated by black arrow) demonstrated supershifting (indicated by red arrow) or disappeared entirely in the presence of LEF1, TCF7 or TCF7L2 antibodies using either MIAPaCa-2 or PANC-1 nuclear extract.

    Journal: Human Molecular Genetics

    Article Title: Functional characterization of a chr13q22.1 pancreatic cancer risk locus reveals long-range interaction and allele-specific effects on DIS3 expression

    doi: 10.1093/hmg/ddw300

    Figure Lengend Snippet: The TCF/LEF family of transcription factors bind the insertion allele of rs386772267 in an allele-specific manner. (A) Electromobility shift assay (EMSA) of rs386772267 deletion and insertion allele probes incubated with PANC-1 nuclear extract with and without unlabelled allelic competitors reveals an allele-specific band for the insertion probe (indicated by the arrow). (B) Alignment of binding sequence motifs for TCF/LEF transcription factors against the insertion probe (insertion allele designated in red). Similarity scores indicate similarity of aligned insertion allele probe to binding sequence matrices for the respective TFs as calculated by Genomatix MatInspector TM . (C and D) The ratios for medium and light labelled proteins bound to ( C ) insertion allele vs. scrambled insertion or ( D ) vs. deletion allele probes. Isotopic labeling for the y-axis experiments were reversed compared to those of the x-axis. Proteins indicated in red (TCF7L2 and SNRPA1) showed consistent preferential binding to the insertion allele vs. both scrambled insertion and deletion allele probes. (E) The insertion probe’s allele-specific EMSA band (indicated by black arrow) demonstrated supershifting (indicated by red arrow) or disappeared entirely in the presence of LEF1, TCF7 or TCF7L2 antibodies using either MIAPaCa-2 or PANC-1 nuclear extract.

    Article Snippet: Supershift experiments were carried out by adding 4 µg anti-LEF1 (sc-8591x, Santa Cruz), anti-TCF7 (sc-271453x, Santa Cruz), anti-TCF7L1 (sc-13026x, Santa Cruz), anti-TCF7L2 (C48H11, Cell Signalling) or anti-IgG (sc-52001, Santa Cruz) antibodies to nuclear extracts for 20 min at room temperature prior to adding labelled probe.

    Techniques: Electro Mobility Shift Assay, Incubation, Binding Assay, Sequencing, Isotopic Labeling

    A – E , Response to mechanical injury in TCF7L2 (transcription factor 7-like 2) overexpressing mice. A , Immunofluorescence (IF) staining of TCF7L2 in wild-type (WT) mouse carotid at baseline and post guidewire injury (relative fluorescent intensities/area and the tunica media (M) area are shown next to the figure. B , TCF7L2 IF intensity in heterozygote overexpressing (TCF7L2-bac) and WT mice before and after injury (relative fluorescent intensities shown underneath). C , Neointima (NI) formation and EVG (elastic tissue fibers-Verhoeff’s Van Gieson) staining in carotid arteries of WT and TCF7L2-bac mice postguidewire injury. Quantifications of NI for both female and male WT and TCF7L2-bac mice postcarotid guidewire injury are shown underneath corresponding figures. The quantification is done by the ratio of intima/M surface area. D and E , Immunofluorescent assessment of SM-MHC (smooth muscle cell myosin heavy chain) and α-SMA (alpha smooth muscle cell actin; both in green) in carotids of TCF7L2-bac, and WT mice, before and after guidewire injury. The relative fluorescent intensities shown underneath. The yellow dotted lined in ( A ) demarcate the NI. *, **, ***, ****Significance with P

    Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

    Article Title: TCF7L2 (Transcription Factor 7-Like 2) Regulation of GATA6 (GATA-Binding Protein 6)-Dependent and -Independent Vascular Smooth Muscle Cell Plasticity and Intimal Hyperplasia

    doi: 10.1161/ATVBAHA.118.311830

    Figure Lengend Snippet: A – E , Response to mechanical injury in TCF7L2 (transcription factor 7-like 2) overexpressing mice. A , Immunofluorescence (IF) staining of TCF7L2 in wild-type (WT) mouse carotid at baseline and post guidewire injury (relative fluorescent intensities/area and the tunica media (M) area are shown next to the figure. B , TCF7L2 IF intensity in heterozygote overexpressing (TCF7L2-bac) and WT mice before and after injury (relative fluorescent intensities shown underneath). C , Neointima (NI) formation and EVG (elastic tissue fibers-Verhoeff’s Van Gieson) staining in carotid arteries of WT and TCF7L2-bac mice postguidewire injury. Quantifications of NI for both female and male WT and TCF7L2-bac mice postcarotid guidewire injury are shown underneath corresponding figures. The quantification is done by the ratio of intima/M surface area. D and E , Immunofluorescent assessment of SM-MHC (smooth muscle cell myosin heavy chain) and α-SMA (alpha smooth muscle cell actin; both in green) in carotids of TCF7L2-bac, and WT mice, before and after guidewire injury. The relative fluorescent intensities shown underneath. The yellow dotted lined in ( A ) demarcate the NI. *, **, ***, ****Significance with P

    Article Snippet: We compared the GATA6 protein levels of WT, TCF7L2 +/− , and TCF7L2-bac mice carotid arteries before and after wire injury by immunofluorescence staining.

    Techniques: Mouse Assay, Immunofluorescence, Staining, BAC Assay

    GATA6 (GATA-binding protein 6) expression, localization, and transcriptional regulation. A , Immunofluorescence (IF) staining of GATA6 in wild-type (WT) and TCF7L2 (transcription factor 7-like 2)-bac mice (mice overexpressing TCF7L2) carotids, before and after guidewire injury (quantification shown next to the figure). B , Subcellular localization of TCF7L2 and GATA6 in tunica media of injured and uninjured carotid artery of WT mice. Colocalization of TCF7L2 and GATA6 in the nucleus and its loss after injury, arrows and arrowheads show GATA6 and TCF7L2, respectively. Nuclei are stained with DAPI (4’,6-diamidino-2-phenylindole). C , TCF7L2, GATA6, and SM-MHC (smooth muscle cell myosin heavy chain) mRNA expression in TCF7L2-bac vs WT vascular smooth muscle cell (VSMC), with and without PDGF-BB (platelet-derived growth factor-BB) stimulation. D , Chromatin immunoprecipitation assay demonstrating TCF7L2 binding to GATA6 promoter in WT and TCF7L2-bac VSMCs; its quantification shown next to the figure. E , GATA6 mRNA expression in WT and TCF7L2 +/− mice aortic lysate, before and after guidewire injury. *, **, and ***Significance with P

    Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

    Article Title: TCF7L2 (Transcription Factor 7-Like 2) Regulation of GATA6 (GATA-Binding Protein 6)-Dependent and -Independent Vascular Smooth Muscle Cell Plasticity and Intimal Hyperplasia

    doi: 10.1161/ATVBAHA.118.311830

    Figure Lengend Snippet: GATA6 (GATA-binding protein 6) expression, localization, and transcriptional regulation. A , Immunofluorescence (IF) staining of GATA6 in wild-type (WT) and TCF7L2 (transcription factor 7-like 2)-bac mice (mice overexpressing TCF7L2) carotids, before and after guidewire injury (quantification shown next to the figure). B , Subcellular localization of TCF7L2 and GATA6 in tunica media of injured and uninjured carotid artery of WT mice. Colocalization of TCF7L2 and GATA6 in the nucleus and its loss after injury, arrows and arrowheads show GATA6 and TCF7L2, respectively. Nuclei are stained with DAPI (4’,6-diamidino-2-phenylindole). C , TCF7L2, GATA6, and SM-MHC (smooth muscle cell myosin heavy chain) mRNA expression in TCF7L2-bac vs WT vascular smooth muscle cell (VSMC), with and without PDGF-BB (platelet-derived growth factor-BB) stimulation. D , Chromatin immunoprecipitation assay demonstrating TCF7L2 binding to GATA6 promoter in WT and TCF7L2-bac VSMCs; its quantification shown next to the figure. E , GATA6 mRNA expression in WT and TCF7L2 +/− mice aortic lysate, before and after guidewire injury. *, **, and ***Significance with P

    Article Snippet: We compared the GATA6 protein levels of WT, TCF7L2 +/− , and TCF7L2-bac mice carotid arteries before and after wire injury by immunofluorescence staining.

    Techniques: Binding Assay, Expressing, Immunofluorescence, Staining, BAC Assay, Mouse Assay, Derivative Assay, Chromatin Immunoprecipitation

    TCF7L2 (transcription factor 7-like 2) stabilizes GATA6 (GATA-binding protein 6) against PDGF (platelet-derived growth factor), forms a complex with it and jointly regulates SM-MHC (smooth muscle cell myosin heavy chain) transcription. A , Time course of GATA6 protein levels after PDGF-BB stimulation in CHX pretreated wild-type (WT) and TCF7L2-bac (TCF7L2 overexpressing) vascular smooth muscle cells (VSMC) by Western blot, quantifications shown next to the figure. B , PDGFRβ (platelet-derived growth factor receptor beta) expression and phosphorylation and activation of its downstream effectors, JNK/SAPK (c-Jun N-terminal kinases/stress-activated kinases) and ERK1/2 (extracellular-signal-regulated kinase 1/2) and in WT and TCF7L2-bac VSMC on PDGF-BB stimulation, quantifications shown next to the figure. C , GATA6 knockdown by siRNA and ( D ) its effect on the protein levels of SM-MHC and SMA (alpha smooth muscle actin) in WT and TCF7L2-bac VSMC, quantifications shown next to the figure. E , Immunoprecipitation of TCF7L2 and GATA6 in WT and TCF7L2-bac VSMC lysates, TCF7L2-specific antibody was used for pulldown, followed by Western blot analysis for GATA6. F , Immunocoprecipitation of TCF7L2 and p300 in WT VSMC lysates; TCF7L2-specific antibody was used for pulldown, followed by Western blot analysis for p300. G and H , Chromatin immunoprecipitation (ChIP) assay demonstrating bindings of TCF7L2 and GATA6 to SM-MHC promoter in VSMC (WT) using ChIP-grade TCF7L2-specific and GATA6-specific antibody, respectively. IgG is used as a negative and input as a positive control. *, **, and ***Significance with P

    Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

    Article Title: TCF7L2 (Transcription Factor 7-Like 2) Regulation of GATA6 (GATA-Binding Protein 6)-Dependent and -Independent Vascular Smooth Muscle Cell Plasticity and Intimal Hyperplasia

    doi: 10.1161/ATVBAHA.118.311830

    Figure Lengend Snippet: TCF7L2 (transcription factor 7-like 2) stabilizes GATA6 (GATA-binding protein 6) against PDGF (platelet-derived growth factor), forms a complex with it and jointly regulates SM-MHC (smooth muscle cell myosin heavy chain) transcription. A , Time course of GATA6 protein levels after PDGF-BB stimulation in CHX pretreated wild-type (WT) and TCF7L2-bac (TCF7L2 overexpressing) vascular smooth muscle cells (VSMC) by Western blot, quantifications shown next to the figure. B , PDGFRβ (platelet-derived growth factor receptor beta) expression and phosphorylation and activation of its downstream effectors, JNK/SAPK (c-Jun N-terminal kinases/stress-activated kinases) and ERK1/2 (extracellular-signal-regulated kinase 1/2) and in WT and TCF7L2-bac VSMC on PDGF-BB stimulation, quantifications shown next to the figure. C , GATA6 knockdown by siRNA and ( D ) its effect on the protein levels of SM-MHC and SMA (alpha smooth muscle actin) in WT and TCF7L2-bac VSMC, quantifications shown next to the figure. E , Immunoprecipitation of TCF7L2 and GATA6 in WT and TCF7L2-bac VSMC lysates, TCF7L2-specific antibody was used for pulldown, followed by Western blot analysis for GATA6. F , Immunocoprecipitation of TCF7L2 and p300 in WT VSMC lysates; TCF7L2-specific antibody was used for pulldown, followed by Western blot analysis for p300. G and H , Chromatin immunoprecipitation (ChIP) assay demonstrating bindings of TCF7L2 and GATA6 to SM-MHC promoter in VSMC (WT) using ChIP-grade TCF7L2-specific and GATA6-specific antibody, respectively. IgG is used as a negative and input as a positive control. *, **, and ***Significance with P

    Article Snippet: We compared the GATA6 protein levels of WT, TCF7L2 +/− , and TCF7L2-bac mice carotid arteries before and after wire injury by immunofluorescence staining.

    Techniques: Binding Assay, Derivative Assay, BAC Assay, Western Blot, Expressing, Activation Assay, Immunoprecipitation, Chromatin Immunoprecipitation, Positive Control

    Cell migration and proliferation in TCF7L2 (transcription factor 7-like 2)-bac (mice overexpressing TCF7L2) and TCF7L2 +/− mice vascular Smooth muscle cell (VSMCs). A , In vitro scratch assay demonstrating slower TCF7L2-bac VSMC migration as compared to wild-type (WT) VSMC (the quantification is shown next to the figure). Area in between the solid black lines indicates the scratch in the VSMC culture. Relative migration in wound-healing assay was assessed as a percentage of the initial gap filled with cells. B and C , Proliferation assay with BrdU (bromodeoxyuridine) incorporation and cell count analysis with and without PDGF-BB (platelet-derived growth factor-BB) stimulation for 24 h in TCF7L2-bac and TCF7L2 +/− VSMC, respectively (quantification shown left underneath). D and E , immunofluorescence staining of carotid artery for P-p53 (phosphorylated TP53) and p16 protein in WT and TCF7L2-bac mice before and after injury (quantification shown underneath D ). F , Protein levels of P-p53, p53, p16, and SM-MHC (smooth muscle cell myosin heavy chain) in WT, TCF7L2-bac and TCF7L2 +/− mice VSMCs by Western blot analysis (quantification shown below, multiple comparison done by ANOVA). G , Relative mRNA expression of p53 in TCF7L2 +/− and TCF7L2-bac mice. *, **, and ***Significance with P

    Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

    Article Title: TCF7L2 (Transcription Factor 7-Like 2) Regulation of GATA6 (GATA-Binding Protein 6)-Dependent and -Independent Vascular Smooth Muscle Cell Plasticity and Intimal Hyperplasia

    doi: 10.1161/ATVBAHA.118.311830

    Figure Lengend Snippet: Cell migration and proliferation in TCF7L2 (transcription factor 7-like 2)-bac (mice overexpressing TCF7L2) and TCF7L2 +/− mice vascular Smooth muscle cell (VSMCs). A , In vitro scratch assay demonstrating slower TCF7L2-bac VSMC migration as compared to wild-type (WT) VSMC (the quantification is shown next to the figure). Area in between the solid black lines indicates the scratch in the VSMC culture. Relative migration in wound-healing assay was assessed as a percentage of the initial gap filled with cells. B and C , Proliferation assay with BrdU (bromodeoxyuridine) incorporation and cell count analysis with and without PDGF-BB (platelet-derived growth factor-BB) stimulation for 24 h in TCF7L2-bac and TCF7L2 +/− VSMC, respectively (quantification shown left underneath). D and E , immunofluorescence staining of carotid artery for P-p53 (phosphorylated TP53) and p16 protein in WT and TCF7L2-bac mice before and after injury (quantification shown underneath D ). F , Protein levels of P-p53, p53, p16, and SM-MHC (smooth muscle cell myosin heavy chain) in WT, TCF7L2-bac and TCF7L2 +/− mice VSMCs by Western blot analysis (quantification shown below, multiple comparison done by ANOVA). G , Relative mRNA expression of p53 in TCF7L2 +/− and TCF7L2-bac mice. *, **, and ***Significance with P

    Article Snippet: We compared the GATA6 protein levels of WT, TCF7L2 +/− , and TCF7L2-bac mice carotid arteries before and after wire injury by immunofluorescence staining.

    Techniques: Migration, BAC Assay, Mouse Assay, In Vitro, Wound Healing Assay, Proliferation Assay, Cell Counting, Derivative Assay, Immunofluorescence, Staining, Western Blot, Expressing

    Response to mechanical injury in TCF7L2 +/− mice. A , Neointima formation and its quantification in carotid arteries of wild-type (WT) and TCF7L2 +/− mice postguidewire injury. B , Immunofluorescence (IF) staining of carotid for TCF7L2 (transcription factor 7-like 2) in uninjured and injured (3 wk after wire injury) TCF7L2 +/− and WT mice. C – D , Immunofluorescent assessment of T α-SMA (alpha smooth muscle cell actin) and SM-MHC (smooth muscle cell myosin heavy chain) in carotids of TCF7L2 +/− and WT mice, before and after guidewire injury. Quantifications are shown next to corresponding figures. *, **, ***Significance with P

    Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

    Article Title: TCF7L2 (Transcription Factor 7-Like 2) Regulation of GATA6 (GATA-Binding Protein 6)-Dependent and -Independent Vascular Smooth Muscle Cell Plasticity and Intimal Hyperplasia

    doi: 10.1161/ATVBAHA.118.311830

    Figure Lengend Snippet: Response to mechanical injury in TCF7L2 +/− mice. A , Neointima formation and its quantification in carotid arteries of wild-type (WT) and TCF7L2 +/− mice postguidewire injury. B , Immunofluorescence (IF) staining of carotid for TCF7L2 (transcription factor 7-like 2) in uninjured and injured (3 wk after wire injury) TCF7L2 +/− and WT mice. C – D , Immunofluorescent assessment of T α-SMA (alpha smooth muscle cell actin) and SM-MHC (smooth muscle cell myosin heavy chain) in carotids of TCF7L2 +/− and WT mice, before and after guidewire injury. Quantifications are shown next to corresponding figures. *, **, ***Significance with P

    Article Snippet: We compared the GATA6 protein levels of WT, TCF7L2 +/− , and TCF7L2-bac mice carotid arteries before and after wire injury by immunofluorescence staining.

    Techniques: Mouse Assay, Immunofluorescence, Staining

    TCF7L2 (transcription factor 7-like 2) overexpression rescues postcarotid injury neointima formation in LRP6 R611C mice. A , Relative intensity of TCF7L2 and GATA6 (GATA-binding protein 6) in LRP6 R611C and LRP6 R611C ; TCF7L2-bac (TCF7L2 overexpressing) mice carotids by immunofluorescence (IF). Magnification (×100) is shown on the right and the area magnified by 5 folds shown on the left . B , Neointima formation ( top row) and IF staining of carotid artery post injury for SM-MHC (smooth muscle cell myosin heavy chain), CD31 ( top row), p-LRP6 (phosphorylated LDLR-related protein 6; second row) and cyclinD1 (third row) in LRP6 R611C vs LRP6 R611C ; TCF7L2-bac mice (nuclei are stained with DAPI [4',6-diamidino-2-phenylindole]); the quantifications for the relative lesion area and SM-MHC are shown. C , TCF7L2 and ( D ) α-SMA (alpha smooth muscle cell actin) IF staining in the aortic roots of the LDLR −/− (low-density lipoprotein receptor knockout) vs wild-type (WT) mice on high-fat diet. Oil Red O staining shown for better visualization of the lesions in the LDLR −/− mice. The boxes denote the sections of aortic roots that are enlarged. The quantifications are shown underneath corresponding figures. *, **, and ***Significance with P

    Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

    Article Title: TCF7L2 (Transcription Factor 7-Like 2) Regulation of GATA6 (GATA-Binding Protein 6)-Dependent and -Independent Vascular Smooth Muscle Cell Plasticity and Intimal Hyperplasia

    doi: 10.1161/ATVBAHA.118.311830

    Figure Lengend Snippet: TCF7L2 (transcription factor 7-like 2) overexpression rescues postcarotid injury neointima formation in LRP6 R611C mice. A , Relative intensity of TCF7L2 and GATA6 (GATA-binding protein 6) in LRP6 R611C and LRP6 R611C ; TCF7L2-bac (TCF7L2 overexpressing) mice carotids by immunofluorescence (IF). Magnification (×100) is shown on the right and the area magnified by 5 folds shown on the left . B , Neointima formation ( top row) and IF staining of carotid artery post injury for SM-MHC (smooth muscle cell myosin heavy chain), CD31 ( top row), p-LRP6 (phosphorylated LDLR-related protein 6; second row) and cyclinD1 (third row) in LRP6 R611C vs LRP6 R611C ; TCF7L2-bac mice (nuclei are stained with DAPI [4',6-diamidino-2-phenylindole]); the quantifications for the relative lesion area and SM-MHC are shown. C , TCF7L2 and ( D ) α-SMA (alpha smooth muscle cell actin) IF staining in the aortic roots of the LDLR −/− (low-density lipoprotein receptor knockout) vs wild-type (WT) mice on high-fat diet. Oil Red O staining shown for better visualization of the lesions in the LDLR −/− mice. The boxes denote the sections of aortic roots that are enlarged. The quantifications are shown underneath corresponding figures. *, **, and ***Significance with P

    Article Snippet: We compared the GATA6 protein levels of WT, TCF7L2 +/− , and TCF7L2-bac mice carotid arteries before and after wire injury by immunofluorescence staining.

    Techniques: Over Expression, Mouse Assay, Binding Assay, BAC Assay, Immunofluorescence, Staining, Knock-Out

    Adiponectin inhibited the expression of β-catenin-associated transcription factor TCF7L2 in pancreatic cancer cells. A and B, The pooled RNA sample from the adiponectin-treated BxPC-3 cells or control cells was subjected to microarray analysis of the mRNA expression profile. A total of 180 genes were identified to be differentially expressed at the mRNA level in response to adiponectin treatment. A, Hierarchical clustering of the 180 differentially expressed genes. The colored images are presented as described; the green and red colors indicate low and high expression levels, respectively. B, GO annotation of differentially expressed genes in BxPC-3 cells induced by adiponectin treatment. The overrepresented GO terms ( P

    Journal: International Journal of Biological Sciences

    Article Title: Adiponectin Suppresses Human Pancreatic Cancer Growth through Attenuating the β-Catenin Signaling Pathway

    doi: 10.7150/ijbs.27420

    Figure Lengend Snippet: Adiponectin inhibited the expression of β-catenin-associated transcription factor TCF7L2 in pancreatic cancer cells. A and B, The pooled RNA sample from the adiponectin-treated BxPC-3 cells or control cells was subjected to microarray analysis of the mRNA expression profile. A total of 180 genes were identified to be differentially expressed at the mRNA level in response to adiponectin treatment. A, Hierarchical clustering of the 180 differentially expressed genes. The colored images are presented as described; the green and red colors indicate low and high expression levels, respectively. B, GO annotation of differentially expressed genes in BxPC-3 cells induced by adiponectin treatment. The overrepresented GO terms ( P

    Article Snippet: Consistent with the mRNA level, the protein level of TCF7L2 was substantially reduced after adiponectin treatment (Fig. D), suggesting that TCF7L2 may participate in the function of adiponectin.

    Techniques: Expressing, Microarray

    Proposed mechanisms of adiponectin-induced growth inhibition of pancreatic cancer. The binding of adiponectin to its receptors AdipoR1 and AdipoR2 on pancreatic cancer cells inhibits the phosphorylation of GSK-3β, which promotes the degradation of β-catenin via enhancing its phosphorylation. Moreover, the activation of adiponectin signaling suppresses the expression of β-catenin-associated transcription factor TCF7L2 in pancreatic cancer cells. Both of these effects lead to the transcriptional inhibition of β-catenin-targeted genes, such as cyclin D1.In consequence, the proliferation of pancreatic cancer cells is inhibited. AdipoQ, adiponectin.

    Journal: International Journal of Biological Sciences

    Article Title: Adiponectin Suppresses Human Pancreatic Cancer Growth through Attenuating the β-Catenin Signaling Pathway

    doi: 10.7150/ijbs.27420

    Figure Lengend Snippet: Proposed mechanisms of adiponectin-induced growth inhibition of pancreatic cancer. The binding of adiponectin to its receptors AdipoR1 and AdipoR2 on pancreatic cancer cells inhibits the phosphorylation of GSK-3β, which promotes the degradation of β-catenin via enhancing its phosphorylation. Moreover, the activation of adiponectin signaling suppresses the expression of β-catenin-associated transcription factor TCF7L2 in pancreatic cancer cells. Both of these effects lead to the transcriptional inhibition of β-catenin-targeted genes, such as cyclin D1.In consequence, the proliferation of pancreatic cancer cells is inhibited. AdipoQ, adiponectin.

    Article Snippet: Consistent with the mRNA level, the protein level of TCF7L2 was substantially reduced after adiponectin treatment (Fig. D), suggesting that TCF7L2 may participate in the function of adiponectin.

    Techniques: Inhibition, Binding Assay, Activation Assay, Expressing

    Distribution of LEF1, TCF7L2, and β-catenin proteins in the adult prethalamus and epithalamus. a TCF7L2 and nuclear β-catenin are present in the subsets of cells in the pregeniculate nucleus. b A clear border is seen between LEF1-, TCF7L2-, and nuclear β-catenin-positive cells between the thalamus and reticular thalamic nucleus. c In the habenula, LEF1, TCF7L2, and nuclear β-catenin are present only in the medial part. DLG dorsal lateral geniculate; LHb lateral habenula; MHb medial habenula; PG pregeniculate nucleus; RT reticular thalamic nucleus; VPL ventral posterolateral thalamic nucleus. Scale bar 100 μm

    Journal: Brain Structure & Function

    Article Title: Postnatal isoform switch and protein localization of LEF1 and TCF7L2 transcription factors in cortical, thalamic, and mesencephalic regions of the adult mouse brain

    doi: 10.1007/s00429-012-0474-6

    Figure Lengend Snippet: Distribution of LEF1, TCF7L2, and β-catenin proteins in the adult prethalamus and epithalamus. a TCF7L2 and nuclear β-catenin are present in the subsets of cells in the pregeniculate nucleus. b A clear border is seen between LEF1-, TCF7L2-, and nuclear β-catenin-positive cells between the thalamus and reticular thalamic nucleus. c In the habenula, LEF1, TCF7L2, and nuclear β-catenin are present only in the medial part. DLG dorsal lateral geniculate; LHb lateral habenula; MHb medial habenula; PG pregeniculate nucleus; RT reticular thalamic nucleus; VPL ventral posterolateral thalamic nucleus. Scale bar 100 μm

    Article Snippet: Antibody characterization Anti-TCF4 (6H5-3) mouse monoclonal antibody was raised against amino acids 31-331 of human TCF7L2 (Millipore).

    Techniques:

    Distribution of LEF1, TCF7L2, and β-catenin proteins in the adult thalamus. a Photomicrographs show the localization of LEF1, TCF7L2, and nuclear β-catenin at different rostro-caudal levels of the thalamus. Double immunofluorescent staining shows that in thalamic cells that express b LEF1 and c TCF7L2 exhibit nuclear localization of β-catenin. d The colabeling with NeuN confirmed the neuronal phenotype of TCF7L2-poitive cells. For abbreviations, see list. Scale bar 200 μm for a . Scale bar 50 μm for b – d

    Journal: Brain Structure & Function

    Article Title: Postnatal isoform switch and protein localization of LEF1 and TCF7L2 transcription factors in cortical, thalamic, and mesencephalic regions of the adult mouse brain

    doi: 10.1007/s00429-012-0474-6

    Figure Lengend Snippet: Distribution of LEF1, TCF7L2, and β-catenin proteins in the adult thalamus. a Photomicrographs show the localization of LEF1, TCF7L2, and nuclear β-catenin at different rostro-caudal levels of the thalamus. Double immunofluorescent staining shows that in thalamic cells that express b LEF1 and c TCF7L2 exhibit nuclear localization of β-catenin. d The colabeling with NeuN confirmed the neuronal phenotype of TCF7L2-poitive cells. For abbreviations, see list. Scale bar 200 μm for a . Scale bar 50 μm for b – d

    Article Snippet: Antibody characterization Anti-TCF4 (6H5-3) mouse monoclonal antibody was raised against amino acids 31-331 of human TCF7L2 (Millipore).

    Techniques: Staining

    Distribution of LEF1, TCF7L2, and β-catenin proteins in the adult pretectum and midbrain. a The pretectal area exhibited TCF7L2- and β-catenin-positive cells, b which have a neuronal phenotype. c In the midbrain, LEF1 and nuclear β-catenin are expressed in all layers of the superior colliculus, whereas some TCF7L2-positive cells are present only in intermediate and deep layers. d The periaqueductal gray showed staining of both transcription factors and nuclear β-catenin in the lateral and dorsal parts. e Many TCF7L2-positive cells and some with nuclear β-catenin are present in the inferior colliculus, and f they exhibit a neuronal phenotype. g TCF7L2 is also expressed in the medial portion of the interpeduncular nucleus. dlPAG dorsolateral periaqueductal gray; dmPAG dorsomedial periaqueductal gray; DpG deep gray layers of the superior colliculus; IC inferior colliculus; InG intermediate gray layer of the superior colliculus; InW intermediate white layer of the superior colliculus; IPN interpeduncular nucleus; lPAG lateral periaqueductal gray; Op optic nerve layer of the superior colliculus; pc posterior commissure; Pn pontine nuclei; PT pretectal region; SuG superficial gray layer of the superior colliculus. Scale bar 100 μm for a . Scale bar 20 μm for b and f . Scale bar 200 μm for c – e

    Journal: Brain Structure & Function

    Article Title: Postnatal isoform switch and protein localization of LEF1 and TCF7L2 transcription factors in cortical, thalamic, and mesencephalic regions of the adult mouse brain

    doi: 10.1007/s00429-012-0474-6

    Figure Lengend Snippet: Distribution of LEF1, TCF7L2, and β-catenin proteins in the adult pretectum and midbrain. a The pretectal area exhibited TCF7L2- and β-catenin-positive cells, b which have a neuronal phenotype. c In the midbrain, LEF1 and nuclear β-catenin are expressed in all layers of the superior colliculus, whereas some TCF7L2-positive cells are present only in intermediate and deep layers. d The periaqueductal gray showed staining of both transcription factors and nuclear β-catenin in the lateral and dorsal parts. e Many TCF7L2-positive cells and some with nuclear β-catenin are present in the inferior colliculus, and f they exhibit a neuronal phenotype. g TCF7L2 is also expressed in the medial portion of the interpeduncular nucleus. dlPAG dorsolateral periaqueductal gray; dmPAG dorsomedial periaqueductal gray; DpG deep gray layers of the superior colliculus; IC inferior colliculus; InG intermediate gray layer of the superior colliculus; InW intermediate white layer of the superior colliculus; IPN interpeduncular nucleus; lPAG lateral periaqueductal gray; Op optic nerve layer of the superior colliculus; pc posterior commissure; Pn pontine nuclei; PT pretectal region; SuG superficial gray layer of the superior colliculus. Scale bar 100 μm for a . Scale bar 20 μm for b and f . Scale bar 200 μm for c – e

    Article Snippet: Antibody characterization Anti-TCF4 (6H5-3) mouse monoclonal antibody was raised against amino acids 31-331 of human TCF7L2 (Millipore).

    Techniques: Staining

    Expression analysis of the Lef1 , Tcf7, Tcf7l1, and Tcf7l2 genes. mRNA copy numbers of a Lef1 , b Tcf7 , c Tcf7l1 , and d Tcf7l2 were measured in the developing and adult cortex, thalamus, and midbrain and adult thymus and colon. The results are shown as mRNA copy number per 1 ng of total RNA (estimated to correspond to ~40 brain cells). Copy numbers were calculated using standard curves generated with serial dilutions of linearized plasmids that carry Lef1 , Tcf7 , Tcf7l1 , and Tcf7l2 cDNAs. The data are expressed as mean ± SD ( n = 3), except for the thymus and colon. Comparisons between copy numbers of full-length (FL) and truncated isoforms of e Lef1 and f Tcf7l2 in the developing thalamus were made using different primer pairs, the positions of which are marked in Figs. 2 a and 3 a, respectively. The data are expressed as mean ± SD ( n = 3). g The pie chart indicates the percentage contribution of full-length ( FL ) and dominant-negative ( dn ) Tcf7l2 isoforms in the embryonic and adult thalamus

    Journal: Brain Structure & Function

    Article Title: Postnatal isoform switch and protein localization of LEF1 and TCF7L2 transcription factors in cortical, thalamic, and mesencephalic regions of the adult mouse brain

    doi: 10.1007/s00429-012-0474-6

    Figure Lengend Snippet: Expression analysis of the Lef1 , Tcf7, Tcf7l1, and Tcf7l2 genes. mRNA copy numbers of a Lef1 , b Tcf7 , c Tcf7l1 , and d Tcf7l2 were measured in the developing and adult cortex, thalamus, and midbrain and adult thymus and colon. The results are shown as mRNA copy number per 1 ng of total RNA (estimated to correspond to ~40 brain cells). Copy numbers were calculated using standard curves generated with serial dilutions of linearized plasmids that carry Lef1 , Tcf7 , Tcf7l1 , and Tcf7l2 cDNAs. The data are expressed as mean ± SD ( n = 3), except for the thymus and colon. Comparisons between copy numbers of full-length (FL) and truncated isoforms of e Lef1 and f Tcf7l2 in the developing thalamus were made using different primer pairs, the positions of which are marked in Figs. 2 a and 3 a, respectively. The data are expressed as mean ± SD ( n = 3). g The pie chart indicates the percentage contribution of full-length ( FL ) and dominant-negative ( dn ) Tcf7l2 isoforms in the embryonic and adult thalamus

    Article Snippet: Antibody characterization Anti-TCF4 (6H5-3) mouse monoclonal antibody was raised against amino acids 31-331 of human TCF7L2 (Millipore).

    Techniques: Expressing, Generated, Dominant Negative Mutation

    Distribution of LEF1, TCF7L2, and β-catenin proteins in the telencephalon. a , b LEF1 is highly expressed in the entorhinal cortex and does not co-localize with nuclear β-catenin. c Single TCF7L2 non-neuronal cells are found in the corpus callosum and cerebral cortex. e The expression of TCF7L2 and nuclear β-catenin is found in the substantia innominata and neighboring structures in f neuronal and non-neuronal cells. ac anterior commissure; cc corpus callosum; CX cerebral cortex; Ent entorhinal cortex; HDB nucleus of the horizontal limb of the diagonal band; LPO lateral preoptic area; SI substantia innominata; VP ventral pallidum. Scale bar 200 μm for a and e . Scale bar 25 μm in b , c , d , and f

    Journal: Brain Structure & Function

    Article Title: Postnatal isoform switch and protein localization of LEF1 and TCF7L2 transcription factors in cortical, thalamic, and mesencephalic regions of the adult mouse brain

    doi: 10.1007/s00429-012-0474-6

    Figure Lengend Snippet: Distribution of LEF1, TCF7L2, and β-catenin proteins in the telencephalon. a , b LEF1 is highly expressed in the entorhinal cortex and does not co-localize with nuclear β-catenin. c Single TCF7L2 non-neuronal cells are found in the corpus callosum and cerebral cortex. e The expression of TCF7L2 and nuclear β-catenin is found in the substantia innominata and neighboring structures in f neuronal and non-neuronal cells. ac anterior commissure; cc corpus callosum; CX cerebral cortex; Ent entorhinal cortex; HDB nucleus of the horizontal limb of the diagonal band; LPO lateral preoptic area; SI substantia innominata; VP ventral pallidum. Scale bar 200 μm for a and e . Scale bar 25 μm in b , c , d , and f

    Article Snippet: Antibody characterization Anti-TCF4 (6H5-3) mouse monoclonal antibody was raised against amino acids 31-331 of human TCF7L2 (Millipore).

    Techniques: Expressing

    Analysis of alternative splicing of Tcf7l2 genes in the mouse thalamus. a Representative intron–exon structure of the Tcf7l2 gene. Long introns are represented by a double slash . Blue boxes indicate the alternatively spliced exons. Important protein domains are marked by red , green and blue lines under the exons by which there are encoded. The position and orientation of the primers used for RT-PCR are indicated by horizontal arrows . The horizontal lines with “Tcf7l2 FL” and “Tcf7l2” indicate the regions of the gene that were tested in the RT-qPCR analysis in Fig. 1 . b RT-PCR expression analysis of alternative splicing of exon 4. c RT-PCR expression analysis of alternative splicing of exon 5. d RT-PCR expression analysis of alternative splicing of exons 13–16. Colon transcript resulted from inclusion of exon 14 or 15 translated from different alternative reading frame ( ARF ). e Western blot analysis of the endogenous TCF7L2 protein in the E18.5 and adult cortex ( Cx ), thalamus ( Th ), and midbrain ( Mb ) and adult colon. As a control, HeLa cells transfected with TCF7L2-E2 and TCF7L2-S3 expression plasmids were used

    Journal: Brain Structure & Function

    Article Title: Postnatal isoform switch and protein localization of LEF1 and TCF7L2 transcription factors in cortical, thalamic, and mesencephalic regions of the adult mouse brain

    doi: 10.1007/s00429-012-0474-6

    Figure Lengend Snippet: Analysis of alternative splicing of Tcf7l2 genes in the mouse thalamus. a Representative intron–exon structure of the Tcf7l2 gene. Long introns are represented by a double slash . Blue boxes indicate the alternatively spliced exons. Important protein domains are marked by red , green and blue lines under the exons by which there are encoded. The position and orientation of the primers used for RT-PCR are indicated by horizontal arrows . The horizontal lines with “Tcf7l2 FL” and “Tcf7l2” indicate the regions of the gene that were tested in the RT-qPCR analysis in Fig. 1 . b RT-PCR expression analysis of alternative splicing of exon 4. c RT-PCR expression analysis of alternative splicing of exon 5. d RT-PCR expression analysis of alternative splicing of exons 13–16. Colon transcript resulted from inclusion of exon 14 or 15 translated from different alternative reading frame ( ARF ). e Western blot analysis of the endogenous TCF7L2 protein in the E18.5 and adult cortex ( Cx ), thalamus ( Th ), and midbrain ( Mb ) and adult colon. As a control, HeLa cells transfected with TCF7L2-E2 and TCF7L2-S3 expression plasmids were used

    Article Snippet: Antibody characterization Anti-TCF4 (6H5-3) mouse monoclonal antibody was raised against amino acids 31-331 of human TCF7L2 (Millipore).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Expressing, Western Blot, Transfection

    TCF7L2 protein expression . Western Blot showing lower amounts of TCF7L2 protein expression in MDA-MB-231 EpCAM cells compared to their empty vector counterparts.

    Journal: BMC Cancer

    Article Title: Effects of EpCAM overexpression on human breast cancer cell lines

    doi: 10.1186/1471-2407-11-45

    Figure Lengend Snippet: TCF7L2 protein expression . Western Blot showing lower amounts of TCF7L2 protein expression in MDA-MB-231 EpCAM cells compared to their empty vector counterparts.

    Article Snippet: Antibodies used for Western analysis were: C-10 (mouse monoclonal against amino acids 24-93 of human EpCAM, Santa Cruz Biotechnology), E144 (rabbit monoclonal antibody against the C-terminus of human EpCAM, Epitomics), β-Catenin (BD Biosciences), pan-actin (Ab-5 Clone, ACTN05, Neomarkers), GAPDH 6C5 (Santa Cruz Biotechnology), lamin A/C (2032, Cell Signalling) and anti-TCF7L2 (clone 6H5-3, Upstate Biotechnology).

    Techniques: Expressing, Western Blot, Multiple Displacement Amplification, Plasmid Preparation

    Schematic representation of TCF7L2, SFRP1 and ITF-2 . (A) TCF7L2 can be divided in 3 major domains: the β-catenin binding domain (CTNNB1), the DNA-binding HMG box and a C-terminal domain which is variable in many isoforms. Only the long form is able to bind to the COOH-terminal binding protein (CtPB), the C-terminus has an important regulatory function. The presence of the CtBP motif is required for the possible repressor function of TCF7L2. Many splicing variants ( > 16) have been described and each cell expresses more than one variant [ 32 , 37 ]. (B) SFRP1 is a secreted protein and contains a frizzled typical cysteine rich domain (Fz-CRD) which displays the putative Wnt-ligand binding site. The C-terminal part of the protein shows homology to netrin (NTR), an extracelluar protein involved in axonal guidance. NTR domains are involved in protein binding and contain segments rich of positively charged residues. This region is reported to bind to heparin [ 34 ]. (C) ITF-2 binds to DNA via its basic-helix-loop-helix (bHLH) domain and can be regulated by β-catenin. The Pitt-Hopkins syndrome (PTHS), a rare syndromic encephalopathy, is caused by ITF-2 haploinsufficency. At least 3 isoformal sequences have been described. The two major isoforms, ITF-2A and ITF-2B, have different N-termini resulting from alternative promoter usage [ 30 ]. (⋆ = glycosilation, p = phosphorylation. Swissprot accession numbers Q9NQB0 , Q8N474 , P15884 )

    Journal: BMC Cancer

    Article Title: Effects of EpCAM overexpression on human breast cancer cell lines

    doi: 10.1186/1471-2407-11-45

    Figure Lengend Snippet: Schematic representation of TCF7L2, SFRP1 and ITF-2 . (A) TCF7L2 can be divided in 3 major domains: the β-catenin binding domain (CTNNB1), the DNA-binding HMG box and a C-terminal domain which is variable in many isoforms. Only the long form is able to bind to the COOH-terminal binding protein (CtPB), the C-terminus has an important regulatory function. The presence of the CtBP motif is required for the possible repressor function of TCF7L2. Many splicing variants ( > 16) have been described and each cell expresses more than one variant [ 32 , 37 ]. (B) SFRP1 is a secreted protein and contains a frizzled typical cysteine rich domain (Fz-CRD) which displays the putative Wnt-ligand binding site. The C-terminal part of the protein shows homology to netrin (NTR), an extracelluar protein involved in axonal guidance. NTR domains are involved in protein binding and contain segments rich of positively charged residues. This region is reported to bind to heparin [ 34 ]. (C) ITF-2 binds to DNA via its basic-helix-loop-helix (bHLH) domain and can be regulated by β-catenin. The Pitt-Hopkins syndrome (PTHS), a rare syndromic encephalopathy, is caused by ITF-2 haploinsufficency. At least 3 isoformal sequences have been described. The two major isoforms, ITF-2A and ITF-2B, have different N-termini resulting from alternative promoter usage [ 30 ]. (⋆ = glycosilation, p = phosphorylation. Swissprot accession numbers Q9NQB0 , Q8N474 , P15884 )

    Article Snippet: Antibodies used for Western analysis were: C-10 (mouse monoclonal against amino acids 24-93 of human EpCAM, Santa Cruz Biotechnology), E144 (rabbit monoclonal antibody against the C-terminus of human EpCAM, Epitomics), β-Catenin (BD Biosciences), pan-actin (Ab-5 Clone, ACTN05, Neomarkers), GAPDH 6C5 (Santa Cruz Biotechnology), lamin A/C (2032, Cell Signalling) and anti-TCF7L2 (clone 6H5-3, Upstate Biotechnology).

    Techniques: Binding Assay, Variant Assay, Ligand Binding Assay, Protein Binding

    Generation of pancreas-specific Tcf7l2 null mice. a) Schematic showing wild-type (top) and conditional inactivation (bottom) of the Tcf7l2 allelle with binding sites for primers used for genotyping (dotted lines). Black boxes represent exons with exon number indicated in white. b) Immunohistochemical analysis of pancreata for Tcf7l2 and insulin. Scale bar = 20 micrometre. c) Real time PCR analysis of gene expression in islets of pTcf7l2 mice (black bar) and wild-type littermate control (dotted/grey bar). Tcf7l2 gene expression was undetected (UD; Ct > 40) in pTcf7l2 mice. Ccnd1, cyclin D1 gene.

    Journal: Diabetologia

    Article Title: Abnormal glucose tolerance and insulin secretion in pancreas-specific Tcf7l2 null mice

    doi: 10.1007/s00125-012-2600-7

    Figure Lengend Snippet: Generation of pancreas-specific Tcf7l2 null mice. a) Schematic showing wild-type (top) and conditional inactivation (bottom) of the Tcf7l2 allelle with binding sites for primers used for genotyping (dotted lines). Black boxes represent exons with exon number indicated in white. b) Immunohistochemical analysis of pancreata for Tcf7l2 and insulin. Scale bar = 20 micrometre. c) Real time PCR analysis of gene expression in islets of pTcf7l2 mice (black bar) and wild-type littermate control (dotted/grey bar). Tcf7l2 gene expression was undetected (UD; Ct > 40) in pTcf7l2 mice. Ccnd1, cyclin D1 gene.

    Article Snippet: Mice carrying conditional knockout alleles of Tcf7l2 ( Tcf7l2 -flox) in which exon 1 was flanked by Lox P sites (Genoway, Lyon, Fr) were backcrossed for five generations into a C57BL/6 background.

    Techniques: Mouse Assay, Binding Assay, Immunohistochemistry, Real-time Polymerase Chain Reaction, Expressing