80c7b3adad54a2602d39a97595dfbbce pmc_29379130.pdf 7abe77ca489d0cec79b7433a432e3abd01122164 pmc_29379130.pdf 169a6fc545f242f14efa672de34d08f8ab3cca143a1f153f9a18c4c2ebe3319c pmc_29379130.pdf Title: DUSP5 is methylated in CIMP-high colorectal cancer but is not a major regulator of intestinal cell proliferation and tumorigenesis Subject: Scientific Reports, doi:10.1038/s41598-018-20176-9 Keywords: Author: Lars Tamp#x000F6;gel Creator: Springer Producer: iText® 5.3.5 ©2000-2012 1T3XT BVBA (AGPL-version) CreationDate: Thu Jan 18 19:53:39 2018 CET ModDate: Tue Jan 23 14:07:21 2018 CET Tagged: no UserProperties: no Suspects: no Form: AcroForm JavaScript: no Pages: 11 Encrypted: no Page size: 595.276 x 782.362 pts Page rot: 0 File size: 7286189 bytes Optimized: no PDF version: 1.4 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- LZBDEM+Corbel-Bold CID TrueType Identity-H yes yes yes 69 0 YCUSEM+Corbel CID TrueType Identity-H yes yes yes 416 0 JINSCQ+TimesNewRomanPSMT Type 1C WinAnsi yes yes yes 417 0 LKDNLX+Corbel-Bold2 Type 1C WinAnsi yes yes yes 71 0 XDSITI+Corbel2 Type 1C WinAnsi yes yes yes 73 0 QJOHWC+Corbel-BoldItalic CID TrueType Identity-H yes yes yes 74 0 MinionPro-Regular Type 1C Custom yes no yes 75 0 LKDNLX+Corbel-Bold2 Type 1C WinAnsi yes yes yes 332 0 XDSITI+Corbel2 Type 1C WinAnsi yes yes yes 333 0 YCUSEM+Corbel CID TrueType Identity-H yes yes yes 453 0 MyriadPro-It Type 1C WinAnsi yes no yes 335 0 MinionPro-It Type 1C Custom yes no yes 70 0 EuclidSymbol Type 1C Custom yes no yes 72 0 XDSITI+Corbel2 Type 1C WinAnsi yes yes yes 327 0 YCUSEM+Corbel CID TrueType Identity-H yes yes yes 334 0 MinionPro-Bold Type 1C WinAnsi yes no yes 106 0 XDSITI+Corbel2 Type 1C WinAnsi yes yes yes 345 0 YCUSEM+Corbel CID TrueType Identity-H yes yes yes 347 0 PTWYDE+STIXGeneral-Regular Type 1C Custom yes yes yes 122 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2022-06-14 02:29:46 CEST RepresentationInformation: pmc_29379130.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2018-03-05 03:49:04 CET Size: 7286189 Format: PDF Version: 1.4 Status: Well-Formed, but not valid SignatureMatches: PDF-hul ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 1. Downregulation of DUSP5 mRNA expression in CRC. DUSP5 mRNA expression in normal colonic mucosa and colorectal tumours determined by analysis of (A) an in-house Affymetrix microarray dataset and (B) RNA-Seq data available from t:46' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 1. Downregulation of DUSP5 mRNA expression in CRC. DUSP5 mRNA expression in normal colonic mucosa and colorectal tumours determined by analysis of (A) an in-house Affymetrix microarray dataset and (B) RNA-Seq data available from :47' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 1. Downregulation of DUSP5 mRNA expression in CRC. DUSP5 mRNA expression in normal colonic mucosa and colorectal tumours determined by analysis of (A) an in-house Affymetrix microarray dataset and (B) RNA-Seq data available from:48' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 1. Downregulation of DUSP5 mRNA expression in CRC. DUSP5 mRNA expression in normal colonic mucosa and colorectal tumours determined by analysis of (A) an in-house Affymetrix microarray dataset and (B) RNA-Seq data available fro:49' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 2. Induction of DUSP5 by EGFR-ERK signalling in CRC cells. Time-course showing EGF-mediated induction of (A) p-ERK1/2 and DUSP5 protein expression, and (B) DUSP5 and c-FOS gene expression in LIM1215 cells. (C) Induction of DU:52' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 2. Induction of DUSP5 by EGFR-ERK signalling in CRC cells. Time-course showing EGF-mediated induction of (A) p-ERK1/2 and DUSP5 protein expression, and (B) DUSP5 and c-FOS gene expression in LIM1215 cells. (C) Induction of D:53' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 2. Induction of DUSP5 by EGFR-ERK signalling in CRC cells. Time-course showing EGF-mediated induction of (A) p-ERK1/2 and DUSP5 protein expression, and (B) DUSP5 and c-FOS gene expression in LIM1215 cells. (C) Induction of :54' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 3. DUSP5 promoter methylation in CRC cell lines and tumours. (A) Schematic of the DUSP5 gene showing the location of the CpG island (highlighted in green). The transcription start site is indicated by dotted arrow. Zoomed image show:56' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 2. Induction of DUSP5 by EGFR-ERK signalling in CRC cells. Time-course showing EGF-mediated induction of (A) p-ERK1/2 and DUSP5 protein expression, and (B) DUSP5 and c-FOS gene expression in LIM1215 cells. (C) Induction of :57' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 3. DUSP5 promoter methylation in CRC cell lines and tumours. (A) Schematic of the DUSP5 gene showing the location of the CpG island (highlighted in green). The transcription start site is indicated by dotted arrow. Zoomed image sh:59' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 3. DUSP5 promoter methylation in CRC cell lines and tumours. (A) Schematic of the DUSP5 gene showing the location of the CpG island (highlighted in green). The transcription start site is indicated by dotted arrow. Zoomed image s:60' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 3. DUSP5 promoter methylation in CRC cell lines and tumours. (A) Schematic of the DUSP5 gene showing the location of the CpG island (highlighted in green). The transcription start site is indicated by dotted arrow. Zoomed image :61' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 4. DUSP5 regulation of basal ERK signaling in CRC cells. (A) Relationship between DUSP5 expression and levels of phosphorylated and total ERK1/2 in nuclear extracts of CRC cell lines. HDAC3 was used as loading control. (B) Analysis of:65' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 4. DUSP5 regulation of basal ERK signaling in CRC cells. (A) Relationship between DUSP5 expression and levels of phosphorylated and total ERK1/2 in nuclear extracts of CRC cell lines. HDAC3 was used as loading control. (B) Analysis :70' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 4. DUSP5 regulation of basal ERK signaling in CRC cells. (A) Relationship between DUSP5 expression and levels of phosphorylated and total ERK1/2 in nuclear extracts of CRC cell lines. HDAC3 was used as loading control. (B) Analysis:73' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 4. DUSP5 regulation of basal ERK signaling in CRC cells. (A) Relationship between DUSP5 expression and levels of phosphorylated and total ERK1/2 in nuclear extracts of CRC cell lines. HDAC3 was used as loading control. (B) Analysi:74' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 5. DUSP5 regulation of EGF-induced ERK signalling in CRC cells. Effect of DUSP5 knockdown on induction of (A) ERK1/2 phosphorylation, and mRNA levels of (B) DUSP5, (C) FOS, and (D) FOSL1 following treatment of LIM1215 cell w:75' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 6. Impact of DUSP5 overexpression on ERK signaling in the mouse intestine. (A) Expression of human DUSP5 protein in the intestinal epithelium detected by blotting for the Myc-tag. (B) Expression of human DUSP5 mRNA specifically in:78' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 6. Impact of DUSP5 overexpression on ERK signaling in the mouse intestine. (A) Expression of human DUSP5 protein in the intestinal epithelium detected by blotting for the Myc-tag. (B) Expression of human DUSP5 mRNA specifically :81' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 6. Impact of DUSP5 overexpression on ERK signaling in the mouse intestine. (A) Expression of human DUSP5 protein in the intestinal epithelium detected by blotting for the Myc-tag. (B) Expression of human DUSP5 mRNA specifically:82' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 7. Impact of DUSP5 overexpression on intestinal phenotype. Effect of DUSP5 overexpression on cell differentiation as determined by staining for (A) Goblet cell (Alcian Blue) and (B) Paneth cell (lysozyme) markers, and measuring mRNA l:85' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 7. Impact of DUSP5 overexpression on intestinal phenotype. Effect of DUSP5 overexpression on cell differentiation as determined by staining for (A) Goblet cell (Alcian Blue) and (B) Paneth cell (lysozyme) markers, and measuring mRNA:87' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 7. Impact of DUSP5 overexpression on intestinal phenotype. Effect of DUSP5 overexpression on cell differentiation as determined by staining for (A) Goblet cell (Alcian Blue) and (B) Paneth cell (lysozyme) markers, and measuring mRN:88' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 7. Impact of DUSP5 overexpression on intestinal phenotype. Effect of DUSP5 overexpression on cell differentiation as determined by staining for (A) Goblet cell (Alcian Blue) and (B) Paneth cell (lysozyme) markers, and measuring mR:89' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 8. Impact of DUSP5 overexpression on intestinal adenoma formation. Effect of DUSP5 overexpression on the (A) number and (B) size of intestinal tumours that develop upon Apc loss. (C) DUSP5 overexpression does not alter the pattern:95' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd:Figure 8. Impact of DUSP5 overexpression on intestinal adenoma formation. Effect of DUSP5 overexpression on the (A) number and (B) size of intestinal tumours that develop upon Apc loss. (C) DUSP5 overexpression does not alter the patte:97' cannot be found Offset: 6903608 ID: PDF-HUL-122 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'SpringerNature_SciRep_20176.indd: 8. Mandl, M., Slack, D. N. & Keyse, S. M. Specific inactivation and nuclear anchoring of extracellular signal-regulated kinase 2 by the inducible dual-specificity protein phosphatase DUSP5. Mol Cell Biol 25, 1830–1845, https://doi.org/:110' cannot be found Offset: 6903608 MIMEtype: application/pdf PDFMetadata: Objects: 511 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: false CenterWindow: false DisplayDocTitle: false NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseOutlines Language: EN Outlines: Item: Title: DUSP5 is methylated in CIMP-high colorectal cancer but is not a major regulator of intestinal cell proliferation and tumori ... Children: Item: Title: Results Children: Item: Title: DUSP5 is downregulated in colorectal cancers (CRCs). Item: Title: DUSP5 is induced by ERK signalling in CRC cell lines. Item: Title: The DUSP5 promoter is methylated in a subset of CRCs. Item: Title: DUSP5 is not a major regulator of ERK signalling in CRC cells. Item: Title: DUSP5 overexpression in the mouse intestinal epithelium does not alter ERK signalling, intestinal homeostasis or adenoma fo ... Item: Title: Discussion Item: Title: Methods Children: Item: Title: Cell lines and treatments. Item: Title: Quantitative real-time PCR. Item: Title: Bisulfite sequencing. Item: Title: High-resolution melting methylation analysis. Item: Title: Western blotting. Item: Title: Generation of Vil1-DUSP5Tg mice. Item: Title: Immunohistochemistry. Item: Title: Statistical methods. Item: Title: Data availability. Item: Title: Acknowledgements Item: Title: Figure 1 Downregulation of DUSP5 mRNA expression in CRC. Item: Title: Figure 2 Induction of DUSP5 by EGFR-ERK signalling in CRC cells. Item: Title: Figure 3 DUSP5 promoter methylation in CRC cell lines and tumours. Item: Title: Figure 4 DUSP5 regulation of basal ERK signaling in CRC cells. Item: Title: Figure 5 DUSP5 regulation of EGF-induced ERK signalling in CRC cells. Item: Title: Figure 6 Impact of DUSP5 overexpression on ERK signaling in the mouse intestine. Item: Title: Figure 7 Impact of DUSP5 overexpression on intestinal phenotype. Item: Title: Figure 8 Impact of DUSP5 overexpression on intestinal adenoma formation. 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Champ#x000FC;eh Sheren Al-Obaidi Ian Luk Mercedes Damp#x000E1;valos-Salas Fiona Chionh Carmel Murone Daniel D. Buchanan Zac Chatterton Oliver M. Sieber Diego Arango Niall C. Tebbutt David Williams Amardeep S. Dhillon John M. Mariadason 10.1038/s41598-018-20176-9 http://dx.doi.org/10.1038/s41598-018-20176-9 journal Scientific Reports © 2018, The Author(s) 2045-2322 10.1038/s41598-018-20176-9 Springer 2018-01-23T14:07:21+01:00 2018-01-19T00:23:39+05:30 2018-01-23T14:07:21+01:00 True iText® 5.3.5 ©2000-2012 1T3XT BVBA (AGPL-version) VoR uuid:54fad06f-4c60-45d4-acce-449d4d9aef99 uuid:9b6d6cb8-93e8-4909-90aa-dfb31031b855 default 1 converted uuid:217cd558-04cb-4ddf-89ec-f5e5c4d38c9a converted to PDF/A-2b pdfToolbox 2018-01-19T00:24:17+05:30 2 B http://ns.adobe.com/xap/1.0/mm/ xmpMM XMP Media Management Schema internal UUID based identifier for specific incarnation of a document InstanceID URI internal The common identifier for all versions and renditions of a document. OriginalDocumentID URI internal A reference to the original document from which this one is derived. It is a minimal reference; missing components can be assumed to be unchanged. For example, a new version might only need to specify the instance ID and version number of the previous version, or a rendition might only need to specify the instance ID and rendition class of the original. DerivedFrom ResourceRef Identifies a portion of a document. This can be a position at which the document has been changed since the most recent event history (stEvt:changed). For a resource within an xmpMM:Ingredients list, the ResourceRef uses this type to identify both the portion of the containing document that refers to the resource, and the portion of the referenced resource that is referenced. http://ns.adobe.com/xap/1.0/sType/Part# stPart Part http://ns.adobe.com/pdf/1.3/ pdf Adobe PDF Schema internal A name object indicating whether the document has been modified to include trapping information Trapped Text http://ns.adobe.com/pdfx/1.3/ pdfx pdfx internal ID of PDF/X standard GTS_PDFXVersion Text internal Conformance level of PDF/X standard GTS_PDFXConformance Text internal Company creating the PDF Company Text internal Date when document was last modified SourceModified Text internal Mirrors crossmark:DOI doi Text http://www.aiim.org/pdfa/ns/id/ pdfaid PDF/A ID Schema internal Part of PDF/A standard part Integer internal Amendment of PDF/A standard amd Text internal Conformance level of PDF/A standard conformance Text http://prismstandard.org/namespaces/basic/2.0/ prism Prism external The aggregation type specifies the unit of aggregation for a content collection. Comment PRISM recommends that the PRISM Aggregation Type Controlled Vocabulary be used to provide values for this element. Note: PRISM recommends against the use of the #other value currently allowed in this controlled vocabulary. In lieu of using #other please reach out to the PRISM group at info@prismstandard.org to request addition of your term to the Aggregation Type Controlled Vocabulary. aggregationType Text external Copyright copyright Text external The Digital Object Identifier for the article. The DOI may also be used as the dc:identifier. If used as a dc:identifier, the URI form should be captured, and the bare identifier should also be captured using prism:doi. If an alternate unique identifier is used as the required dc:identifier, then the DOI should be specified as a bare identifier within prism:doi only. If the URL associated with a DOI is to be specified, then prism:url may be used in conjunction with prism:doi in order to provide the service endpoint (i.e. the URL). doi Text external ISSN for an electronic version of the issue in which the resource occurs. Permits publishers to include a second ISSN, identifying an electronic version of the issue in which the resource occurs (therefore e(lectronic)Issn. If used, prism:eIssn MUST contain the ISSN of the electronic version. See prism:issn. issn Text external Title of the magazine, or other publication, in which a resource was/will be published. Typically this will be used to provide the name of the magazine an article appeared in as metadata for the article, along with information such as the article title, the publisher, volume, number, and cover date. 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