DOCSLIB.ORG

Effects of Diabetes and Hoxa3 Upon Macrophage Function

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Effects of Diabetes and Hoxa3 Upon Macrophage Function EFFECTS OF DIABETES AND HOXA3 UPON MACROPHAGE FUNCTION A thesis submitted to The University of Manchester for the degree of Doctor of Philosophy Developmental Biology in the Faculty Life Sciences 2015 MATTHEW BURGESS FACULTY OF LIFE SCIENCES List of contents List of contents .............................................................................................................................. 2 List of figures ................................................................................................................................. 7 List of tables .................................................................................................................................. 9 Declaration .................................................................................................................................. 12 Copyright statement ................................................................................................................... 12 Acknowledgement ...................................................................................................................... 13 The author ................................................................................................................................... 14 1 Introduction ........................................................................................................................ 15 1.1 Cutaneous wound healing .......................................................................................... 15 1.1.1 Inflammatory phase ........................................................................................... 17 1.1.2 Inflammation and the chronic wound ................................................................ 19 1.1.3 Proliferation phase .............................................................................................. 19 1.1.4 The proliferation phase and chronic wounds ..................................................... 20 1.1.5 Neovascularisation .............................................................................................. 21 1.1.6 Neovascularisation and chronic wounds ............................................................ 23 1.1.7 Remodelling phase .............................................................................................. 25 1.2 Macrophages .............................................................................................................. 25 1.2.1 Origins of macrophages ...................................................................................... 25 1.2.2 Maturation and activation .................................................................................. 26 1.2.3 Myeloid cells and neovascularisation ................................................................. 28 1.2.4 Macrophages and diabetes ................................................................................. 29 1.3 Endothelial progenitor cells .............................................................................................. 30 1.3.1 Origins of endothelial progenitor cells....................................................................... 30 1.3.2 Isolation of endothelial progenitor cells ............................................................. 31 1.3.3 Roles of endothelial progenitor cells in neovascuarisation ................................ 35 1.4 Transcriptional basis of lineage plasticity ................................................................... 37 1.4.1 Reprogramming in haematopoiesis ........................................................................... 38 1.4.2 The epigenetic landscape – erythroid/monocyte fate........................................ 40 1.5 Hypothesis and Experimental Aims ............................................................................ 41 1.5.1 Investigation of the phenotypic differences between non-diabetic and diabetic macrophages ....................................................................................................................... 41 1) Differentiation of macrophages in models of diabetes .......................................... 41 2) Activation potential of diabetic macrophages ........................................................ 42 3) Interaction of diabetic macrophages with neovascularisation ............................... 42 2 1.5.2 Identification of transcription factors with monocytic to endothelial reprogramming potential.................................................................................................... 43 1) Identification of transcription factors to be tested for endothelial reprogramming potential .......................................................................................................................... 43 2) Development of an assay to screen transcription factors for monocytic to endothelial reprogramming ............................................................................................ 43 3) Assess the effects of the validated transcription factors upon THP-1 cell phenotype in the transdifferentiation assay ..................................................................................... 44 1.5.3 Investigation of the effects of Hoxa3 transcriptional activity upon macrophages 44 1) Effects of Hoxa3 upon macrophage development ................................................. 44 2) Effects of Hoxa3 upon macrophage activation ....................................................... 45 3) Effects of Hoxa3 upon macrophage interactions with neovascularisation ............ 45 2 Materials and methods ....................................................................................................... 46 2.1 Cell culture .................................................................................................................. 46 2.1.1 Cell lines .............................................................................................................. 46 2.1.2 Cell Culture .......................................................................................................... 46 RAW 264.7 cells............................................................................................................... 46 bEnd.5 ............................................................................................................................. 47 L929 cells ......................................................................................................................... 47 THP-1 cells ....................................................................................................................... 47 HUVECs ............................................................................................................................ 48 2.1.3 Primary cell lines ................................................................................................. 48 Murine primary bone marrow cells ................................................................................ 48 Macrophage activation ................................................................................................... 49 2.1.4 Cryostorage of cells ............................................................................................. 50 2.1.5 Nucleofection ...................................................................................................... 50 2.1.6 Conditioned medium culture .............................................................................. 54 Calcium phosphate transfection ..................................................................................... 54 Conditioned medium collection ...................................................................................... 55 Treatment of cells with SP.Hoxa3.mCherry conditioned medium ................................. 55 Pleiotrophin and macrophage colony stimulating factor supplemented medium......... 56 2.1.7 Neovascularisation assays ................................................................................... 56 2.2 PCR .............................................................................................................................. 58 2.2.1 RNA extraction .................................................................................................... 58 2.2.2 DNase treatment ................................................................................................. 58 2.2.3 cDNA generation ................................................................................................. 59 3 2.2.4 Quantitative PCRs ............................................................................................... 59 TaqMan real time PCRs ................................................................................................... 59 SYBR real-time PCRs ........................................................................................................ 59 Relative expression analysis ............................................................................................ 60 2.2.5 Generation of pCRII-TOPO pleiotrophin ............................................................. 64 2.2.6 Sub-cloning of pcDNA3.1/myc-His pleiotrophin ................................................. 65 2.2.7 Colony PCR .......................................................................................................... 66 2.3 Western blotting ......................................................................................................... 66 2.3.1 Reagents .............................................................................................................
Load more

Recommended publications
  • 1714 Gene Comprehensive Cancer Panel Enriched for Clinically Actionable Genes with Additional Biologically Relevant Genes 400-500X Average Coverage on Tumor
    xO GENE PANEL 1714 gene comprehensive cancer panel enriched for clinically actionable genes with additional biologically relevant genes 400-500x average coverage on tumor Genes A-C Genes D-F Genes G-I Genes J-L AATK ATAD2B BTG1 CDH7 CREM DACH1 EPHA1 FES G6PC3 HGF IL18RAP JADE1 LMO1 ABCA1 ATF1 BTG2 CDK1 CRHR1 DACH2 EPHA2 FEV G6PD HIF1A IL1R1 JAK1 LMO2 ABCB1 ATM BTG3 CDK10 CRK DAXX EPHA3 FGF1 GAB1 HIF1AN IL1R2 JAK2 LMO7 ABCB11 ATR BTK CDK11A CRKL DBH EPHA4 FGF10 GAB2 HIST1H1E IL1RAP JAK3 LMTK2 ABCB4 ATRX BTRC CDK11B CRLF2 DCC EPHA5 FGF11 GABPA HIST1H3B IL20RA JARID2 LMTK3 ABCC1 AURKA BUB1 CDK12 CRTC1 DCUN1D1 EPHA6 FGF12 GALNT12 HIST1H4E IL20RB JAZF1 LPHN2 ABCC2 AURKB BUB1B CDK13 CRTC2 DCUN1D2 EPHA7 FGF13 GATA1 HLA-A IL21R JMJD1C LPHN3 ABCG1 AURKC BUB3 CDK14 CRTC3 DDB2 EPHA8 FGF14 GATA2 HLA-B IL22RA1 JMJD4 LPP ABCG2 AXIN1 C11orf30 CDK15 CSF1 DDIT3 EPHB1 FGF16 GATA3 HLF IL22RA2 JMJD6 LRP1B ABI1 AXIN2 CACNA1C CDK16 CSF1R DDR1 EPHB2 FGF17 GATA5 HLTF IL23R JMJD7 LRP5 ABL1 AXL CACNA1S CDK17 CSF2RA DDR2 EPHB3 FGF18 GATA6 HMGA1 IL2RA JMJD8 LRP6 ABL2 B2M CACNB2 CDK18 CSF2RB DDX3X EPHB4 FGF19 GDNF HMGA2 IL2RB JUN LRRK2 ACE BABAM1 CADM2 CDK19 CSF3R DDX5 EPHB6 FGF2 GFI1 HMGCR IL2RG JUNB LSM1 ACSL6 BACH1 CALR CDK2 CSK DDX6 EPOR FGF20 GFI1B HNF1A IL3 JUND LTK ACTA2 BACH2 CAMTA1 CDK20 CSNK1D DEK ERBB2 FGF21 GFRA4 HNF1B IL3RA JUP LYL1 ACTC1 BAG4 CAPRIN2 CDK3 CSNK1E DHFR ERBB3 FGF22 GGCX HNRNPA3 IL4R KAT2A LYN ACVR1 BAI3 CARD10 CDK4 CTCF DHH ERBB4 FGF23 GHR HOXA10 IL5RA KAT2B LZTR1 ACVR1B BAP1 CARD11 CDK5 CTCFL DIAPH1 ERCC1 FGF3 GID4 HOXA11 IL6R KAT5 ACVR2A
    [Show full text]
  • THE ROLE of HOXD10 in the DEVELOPMENT of MOTONEURONS in the POSTERIOR SPINAL CORD by Veeral Shailesh Shah B.S., University of Pi
    THE ROLE OF HOXD10 IN THE DEVELOPMENT OF MOTONEURONS IN THE POSTERIOR SPINAL CORD by Veeral Shailesh Shah B.S., University of Pittsburgh, 2000 Submitted to the Graduate Faculty of University of Pittsburgh School of Medicine Department of Neurobiology in partial fulfillment of the requirements for the degree of Doctor of Philosophy University of Pittsburgh 2006 UNIVERSITY OF PITTSBURGH SCHOOL OF MEDICINE DEPARTMENT OF NEUROBIOLOGY This thesis was presented by Veeral Shailesh Shah It was defended on January 17, 2006 and approved by Paula Monaghan-Nichols, Ph. D, CNUP, Neurobiology Debbie Chapman, Ph. D, Biological Sciences Willi Halfter, Ph. D, CNUP, Neurobiology Carl Lagenaur, Ph. D, CNUP, Neurobiology Cynthia Forehand, Ph. D, University of Vermont, Anatomy and Neurobiology Dissertation Advisor: Cynthia Lance-Jones, Ph. D, CNUP, Neurobiology ii Copyright © by Veeral Shailesh Shah 2007 iii THE ROLE OF HOXD10 IN THE DEVELOPMENT OF MOTONEURONS IN THE POSTERIOR SPINAL CORD Veeral Shah University of Pittsburgh, 2007 Hox genes encode anterior-posterior identity during central nervous system development. Few studies have examined Hox gene function at lumbosacral (LS) levels of the spinal cord, where there is extensive information on normal development. Hoxd10 is expressed at high levels in the embryonic LS spinal cord, but not the thoracic (T) spinal cord. To test the hypothesis that restricted expression of Hoxd10 contributes to the attainment of an LS identity, and specifically an LS motoneuron identity, Hoxd10 was ectopically expressed in T segments in chick embryos via in ovo electroporation. Electroporations were carried out at early neural tube stages (stages 13-15) and at the onset of motoneuron differentiation (stages 17-18).
    [Show full text]
  • PHC1 Maintains Pluripotency by Organizing Genome-Wide Chromatin Interactions of the Nanog Locus
    ARTICLE https://doi.org/10.1038/s41467-021-22871-0 OPEN PHC1 maintains pluripotency by organizing genome-wide chromatin interactions of the Nanog locus Li Chen1,2,3,14, Qiaoqiao Tong1,2,3,14, Xiaowen Chen4,14, Penglei Jiang1,2, Hua Yu1,2, Qianbing Zhao1,2,3, Lingang Sun1,2,3, Chao Liu 1,2,3,5, Bin Gu6, Yuping Zheng 7, Lijiang Fei1,2,3, Xiao Jiang8, Wenjuan Li9,10, Giacomo Volpe 9,10, Mazid MD. Abdul9,10, Guoji Guo 1,2,3, Jin Zhang1,2, Pengxu Qian1,2, Qiming Sun 8, ✉ ✉ ✉ Dante Neculai 7, Miguel A. Esteban9,10,11, Chen Li 12 , Feiqiu Wen4 & Junfeng Ji 1,2,3,13 1234567890():,; Polycomb group (PcG) proteins maintain cell identity by repressing gene expression during development. Surprisingly, emerging studies have recently reported that a number of PcG proteins directly activate gene expression during cell fate determination process. However, the mechanisms by which they direct gene activation in pluripotency remain poorly under- stood. Here, we show that Phc1, a subunit of canonical polycomb repressive complex 1 (cPRC1), can exert its function in pluripotency maintenance via a PRC1-independent activa- tion of Nanog. Ablation of Phc1 reduces the expression of Nanog and overexpression of Nanog partially rescues impaired pluripotency caused by Phc1 depletion. We find that Phc1 interacts with Nanog and activates Nanog transcription by stabilizing the genome-wide chromatin interactions of the Nanog locus. This adds to the already known canonical function of PRC1 in pluripotency maintenance via a PRC1-dependent repression of differentiation genes. Overall, our study reveals a function of Phc1 to activate Nanog transcription through regulating chromatin architecture and proposes a paradigm for PcG proteins to maintain pluripotency.
    [Show full text]
  • A Crucial Role for the Ubiquitously Expressed Transcription Factor Sp1 at Early Stages of Hematopoietic Specification
    This is a repository copy of A crucial role for the ubiquitously expressed transcription factor Sp1 at early stages of hematopoietic specification.. White Rose Research Online URL for this paper: http://eprints.whiterose.ac.uk/83683/ Version: Published Version Article: Gilmour, J, Assi, SA, Jaegle, U et al. (6 more authors) (2014) A crucial role for the ubiquitously expressed transcription factor Sp1 at early stages of hematopoietic specification. Development, 141 (12). 2391 - 2401. ISSN 0950-1991 https://doi.org/10.1242/dev.106054 Reuse Unless indicated otherwise, fulltext items are protected by copyright with all rights reserved. The copyright exception in section 29 of the Copyright, Designs and Patents Act 1988 allows the making of a single copy solely for the purpose of non-commercial research or private study within the limits of fair dealing. The publisher or other rights-holder may allow further reproduction and re-use of this version - refer to the White Rose Research Online record for this item. Where records identify the publisher as the copyright holder, users can verify any specific terms of use on the publisher’s website. Takedown If you consider content in White Rose Research Online to be in breach of UK law, please notify us by emailing [email protected] including the URL of the record and the reason for the withdrawal request. [email protected] https://eprints.whiterose.ac.uk/ © 2014. Published by The Company of Biologists Ltd | Development (2014) 141, 2391-2401 doi:10.1242/dev.106054 RESEARCH ARTICLE STEM CELLS AND REGENERATION A crucial role for the ubiquitously expressed transcription factor Sp1 at early stages of hematopoietic specification Jane Gilmour1, Salam A.
    [Show full text]
  • Supplementary Materials For
    Supplementary Materials for 7q11.23 Syndromes Reveal BAZ1B as a Master Regulator of the Modern Human Face and Validate the Self-Domestication Hypothesis Matteo Zanella, Alessandro Vitriolo, Alejandro Andirko, Pedro Tiago Martins, Stefanie Sturm, Thomas O’Rourke, Magdalena Laugsch, Natascia Malerba, Adrianos Skaros, Sebastiano Trattaro, Pierre-Luc Germain, Giuseppe Merla, Alvaro Rada-Iglesias, Cedric Boeckx and Giuseppe Testa Correspondence to: [email protected]; [email protected] This PDF file includes: Materials and Methods Figs. S1 to S4 Tables S1 to S17 1 Material and Methods Human samples Ethics approvals are reported in the study that established the original iPSC cohort (13) and also apply to the additional samples included in this study (7dupASD3 and CTL4R). Fibroblast reprogramming and iPSC culture WBS1, WBS2, WBS3, WBS4, 7dupASD1, AtWBS1 and CTL2 fibroblasts were reprogrammed using the mRNA Reprogramming kit (Stemgent), while the 7dupASD2 and CTL1R lines were reprogrammed with the microRNA Booster kit (Stemgent). The CTL3 line was reprogrammed by transfection with the STEMCCA polycistronic lentiviral vector followed by Cre-mediated excision of the integrated polycistron. 7dupASD3 and CTL4R fibroblasts were reprogrammed using the SimpliconTM RNA Reprogramming kit (Millipore). Prior to differentiation, iPSC lines were cultured on hESC-qualified Matrigel (BD Biosciences) coated plates, diluted 1:40 in DMEM/F-12 and grown in mTeSR-1 medium (StemCell Technologies). They were passaged upon treatment with Accutase (Sigma) and then plated in mTeSR-1 medium supplemented with 5μM Y-27632 (Sigma). Differentiation Differentiation into NCSCs was performed as previously described (67), with the exception of NCSCs used in the experiment reported in Fig.
    [Show full text]
  • Three-Dimensional Regulation of HOXA Cluster Genes by a Cis-Element in Hematopoietic Stem Cell and Leukemia
    bioRxiv preprint doi: https://doi.org/10.1101/2020.04.16.017533; this version posted April 18, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. Three-dimensional regulation of HOXA cluster genes by a cis-element in hematopoietic stem cell and leukemia. Xue Qing David Wang1, Haley Gore1, Pamela Himadewi1, Fan Feng2, Lu Yang3, Wanding Zhou1,5, Yushuai Liu1, Xinyu Wang4, Chun-wei Chen3, Jianzhong Su4, Jie Liu2, Gerd Pfeifer1,*, Xiaotian Zhang1,* 1. Center for Epigenetics, Van Andel Research Institute, Grand Rapids, Michigan 2. Department of Computational Medicine and Bioinformatics, University of Michigan, Ann Arbor, Michigan. 3. Department of System Biology, City of Hope Cancer Center, California 4. Institute of Biomedical Big Data, Wenzhou Medical University, Wenzhou, China 5. Current address: Children Hospital of Philadelphia, University of Pennsylvania *. These authors jointly supervise this project Correspondence should be addressed to Xiaotian Zhang: [email protected] bioRxiv preprint doi: https://doi.org/10.1101/2020.04.16.017533; this version posted April 18, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. Abstract: Proper gene regulation is crucial for cellular differentiation, and dysregulation of key genes can lead to diseased states such as cancer. The HOX transcription factors play such a role during hematopoiesis, and aberrant expression of certain HOXA genes is found in certain acute myeloid leukemias (AMLs).
    [Show full text]
  • Xo PANEL DNA GENE LIST
    xO PANEL DNA GENE LIST ~1700 gene comprehensive cancer panel enriched for clinically actionable genes with additional biologically relevant genes (at 400 -500x average coverage on tumor) Genes A-C Genes D-F Genes G-I Genes J-L AATK ATAD2B BTG1 CDH7 CREM DACH1 EPHA1 FES G6PC3 HGF IL18RAP JADE1 LMO1 ABCA1 ATF1 BTG2 CDK1 CRHR1 DACH2 EPHA2 FEV G6PD HIF1A IL1R1 JAK1 LMO2 ABCB1 ATM BTG3 CDK10 CRK DAXX EPHA3 FGF1 GAB1 HIF1AN IL1R2 JAK2 LMO7 ABCB11 ATR BTK CDK11A CRKL DBH EPHA4 FGF10 GAB2 HIST1H1E IL1RAP JAK3 LMTK2 ABCB4 ATRX BTRC CDK11B CRLF2 DCC EPHA5 FGF11 GABPA HIST1H3B IL20RA JARID2 LMTK3 ABCC1 AURKA BUB1 CDK12 CRTC1 DCUN1D1 EPHA6 FGF12 GALNT12 HIST1H4E IL20RB JAZF1 LPHN2 ABCC2 AURKB BUB1B CDK13 CRTC2 DCUN1D2 EPHA7 FGF13 GATA1 HLA-A IL21R JMJD1C LPHN3 ABCG1 AURKC BUB3 CDK14 CRTC3 DDB2 EPHA8 FGF14 GATA2 HLA-B IL22RA1 JMJD4 LPP ABCG2 AXIN1 C11orf30 CDK15 CSF1 DDIT3 EPHB1 FGF16 GATA3 HLF IL22RA2 JMJD6 LRP1B ABI1 AXIN2 CACNA1C CDK16 CSF1R DDR1 EPHB2 FGF17 GATA5 HLTF IL23R JMJD7 LRP5 ABL1 AXL CACNA1S CDK17 CSF2RA DDR2 EPHB3 FGF18 GATA6 HMGA1 IL2RA JMJD8 LRP6 ABL2 B2M CACNB2 CDK18 CSF2RB DDX3X EPHB4 FGF19 GDNF HMGA2 IL2RB JUN LRRK2 ACE BABAM1 CADM2 CDK19 CSF3R DDX5 EPHB6 FGF2 GFI1 HMGCR IL2RG JUNB LSM1 ACSL6 BACH1 CALR CDK2 CSK DDX6 EPOR FGF20 GFI1B HNF1A IL3 JUND LTK ACTA2 BACH2 CAMTA1 CDK20 CSNK1D DEK ERBB2 FGF21 GFRA4 HNF1B IL3RA JUP LYL1 ACTC1 BAG4 CAPRIN2 CDK3 CSNK1E DHFR ERBB3 FGF22 GGCX HNRNPA3 IL4R KAT2A LYN ACVR1 BAI3 CARD10 CDK4 CTCF DHH ERBB4 FGF23 GHR HOXA10 IL5RA KAT2B LZTR1 ACVR1B BAP1 CARD11 CDK5 CTCFL DIAPH1 ERCC1 FGF3 GID4 HOXA11
    [Show full text]
  • DNA Methylation Analysis on Purified Neurons and Glia Dissects Age And
    Gasparoni et al. Epigenetics & Chromatin (2018) 11:41 https://doi.org/10.1186/s13072-018-0211-3 Epigenetics & Chromatin RESEARCH Open Access DNA methylation analysis on purifed neurons and glia dissects age and Alzheimer’s disease‑specifc changes in the human cortex Gilles Gasparoni1 , Sebastian Bultmann2, Pavlo Lutsik3, Theo F. J. Kraus4, Sabrina Sordon5, Julia Vlcek4, Vanessa Dietinger4, Martina Steinmaurer4, Melanie Haider4, Christopher B. Mulholland2, Thomas Arzberger4, Sigrun Roeber4, Matthias Riemenschneider5, Hans A. Kretzschmar4, Armin Giese4, Heinrich Leonhardt2 and Jörn Walter1* Abstract Background: Epigenome-wide association studies (EWAS) based on human brain samples allow a deep and direct understanding of epigenetic dysregulation in Alzheimer’s disease (AD). However, strong variation of cell-type propor- tions across brain tissue samples represents a signifcant source of data noise. Here, we report the frst EWAS based on sorted neuronal and non-neuronal (mostly glia) nuclei from postmortem human brain tissues. Results: We show that cell sorting strongly enhances the robust detection of disease-related DNA methylation changes even in a relatively small cohort. We identify numerous genes with cell-type-specifc methylation signatures and document diferential methylation dynamics associated with aging specifcally in neurons such as CLU, SYNJ2 and NCOR2 or in glia RAI1,CXXC5 and INPP5A. Further, we found neuron or glia-specifc associations with AD Braak stage progression at genes such as MCF2L, ANK1, MAP2, LRRC8B, STK32C and S100B. A comparison of our study with previous tissue-based EWAS validates multiple AD-associated DNA methylation signals and additionally specifes their origin to neuron, e.g., HOXA3 or glia (ANK1). In a meta-analysis, we reveal two novel previously unrecognized methylation changes at the key AD risk genes APP and ADAM17.
    [Show full text]
  • Parathyroid Tumor Development Involves Deregulation of Homeobox Genes
    Endocrine-Related Cancer (2008) 15 267–275 Parathyroid tumor development involves deregulation of homeobox genes H-C Jennifer Shen, Jennifer E Rosen, Lauren M Yang, Sharon A Savage3, A Lee Burns1, Carmen M Mateo1, Sunita K Agarwal1, Settara C Chandrasekharappa2, Allen M Spiegel1, Francis S Collins2, Stephen J Marx1 and Steven K Libutti Tumor Angiogenesis Section, Surgery Branch, National Cancer Institute, Building 10, Room 4W-5940, 10 Center Drive, Bethesda, Maryland 20892, USA 1Metabolic Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, Maryland, USA 2Genome Technology Branch, National Human Genome Research Institute, Bethesda, Maryland, USA 3Division of Cancer Epidemiology and Genetics, National Cancer Institute, National Institute of Health, Bethesda, Maryland, USA (Correspondence should be addressed to S K Libutti; Email: [email protected]) A M Spiegel is now at Albert Einstein College of Medicine, Bronx, New York, USA Abstract Multiple endocrine neoplasia type 1 (MEN1) is an autosomal dominant syndrome caused by mutations in the MEN1 tumor suppressor gene. Loss of the functional second copy of the MEN1 gene causes individuals to develop multiple endocrine tumors, primarily affecting the parathyroid, pituitary, and pancreas. While it is clear that the protein encoded by MEN1, menin, suppresses endocrine tumors, its biochemical functions and direct downstream targets remain unclear. Recent studies have suggested that menin may act as a scaffold protein to coordinate gene transcription, and that menin is an oncogenic cofactor for homeobox (HOX) gene expression in hematopoietic cancer. The role of HOX genes in adult cell differentiation is still obscure, but growing evidence suggests that they may play important roles in the development of cancer.
    [Show full text]
  • Papadaki Et Al., 2009 Supplementary
    Papadaki et al., 2009 Supplementary Supplemental Data Index x Supplemental Figures 1-6 x Supplemental Tables 1a, 1b, 2 Papadaki et al., 2009 Supplementary Supplemental Figure 1. Thymocyte restricted inactivation of the Elavl1 locus. + fl (A) Diagrammatic representation of the wild-type (Elavl1P P), floxed (Elavl1P P) and Cre- - recombined (Elavl1P P) Elavl1/HuR loci on mouse chromosome 8; Noted are the loxP sequences (triangles) flanking the selection marker (neo) used in gene targeting and the ATG containing exon 2 (white box); (H) denotes restriction sites for loci mapping. (B) Detection of native (+), targeted (fl) and Cre-recombinant (-) loci in thymocyte DNA extracts from control and test mice following HindIII digestion and Southern blotting. (C) Western blot of total thymic protein extracts probed with ĮHuR Ab + fl/fl indicating the loss of HuR protein in LckCreP PElavl1P P thymi. Į-actin is shown for quantitation. (D) Flow cytometric detection of intracellular mHuR protein in + fl/+ LckCreP PElavl1P P thymocytes (open histogram), and its respective loss in + fl/fl LckCreP PElavl1P P thymocytes (shaded histogram). The dotted histogram depicts the + isotype-matched background staining. (E) Flow cytometric detection of HuRP P or - + + + fl/+ HuRP P cells in gated splenic CD4P Por CD8P P T-cells from 8 week old LckCreP PElavl1P + fl/fl - Pand LckCreP PElavl1P P mice respectively. (F) Enumeration of HuRP P cells in + fl/fl LckCreP PElavl1P P thymocyte subsets and splenic T-cells; Data are percentages (+SEM) derived from the flow cytometric detection of HuR- cells in CD4/CD8/DP and DN gated populations (n=12-15) at 8-10 weeks of age.
    [Show full text]
  • A Study of the Differential Expression Profiles of Keshan Disease Lncrna/Mrna Genes Based on RNA-Seq
    421 Original Article A study of the differential expression profiles of Keshan disease lncRNA/mRNA genes based on RNA-seq Guangyong Huang1, Jingwen Liu2, Yuehai Wang1, Youzhang Xiang3 1Department of Cardiology, Liaocheng People’s Hospital of Shandong University, Liaocheng, China; 2School of Nursing, Liaocheng Vocational & Technical College, Liaocheng, China; 3Shandong Institute for Endemic Disease Control, Jinan, China Contributions: (I) Conception and design: G Huang, Y Xiang; (II) Administrative support: G Huang, Y Wang; (III) Provision of study materials or patients: G Huang, J Liu, Y Xiang; (IV) Collection and assembly of data: G Huang, J Liu; (V) Data analysis and interpretation: J Liu, Y Wang; (VI) Manuscript writing: All authors; (VII) Final approval of manuscript: All authors. Correspondence to: Guangyong Huang, MD, PhD. Department of Cardiology, Liaocheng People’s Hospital of Shandong University, No. 67 of Dongchang Street, Liaocheng 252000, China. Email: [email protected]. Background: This study aims to analyze the differential expression profiles of lncRNA in Keshan disease (KSD) and to explore the molecular mechanism of the disease occurrence and development. Methods: RNA-seq technology was used to construct the lncRNA/mRNA expression library of a KSD group (n=10) and a control group (n=10), and then Cuffdiff software was used to obtain the gene lncRNA/ mRNA FPKM value as the expression profile of lncRNA/mRNA. The fold changes between the two sets of samples were calculated to obtain differential lncRNA/mRNA expression profiles, and a bioinformatics analysis of differentially expressed genes was performed. Results: A total of 89,905 lncRNAs and 20,315 mRNAs were detected.
    [Show full text]
  • Multiple Roles for HOXA3 in Regulating Thymus and Parathyroid Differentiation and Morphogenesis in Mouse Jena L
    University of South Carolina Scholar Commons Faculty Publications Department of Natural Sciences 2014 Multiple Roles for HOXA3 in Regulating Thymus and Parathyroid Differentiation and Morphogenesis in Mouse Jena L. Chojnowski University of South Carolina - Beaufort, [email protected] Kyoko Masuda University of Georgia Heidi A. Trau University of Georgia Kirk Thomas University of Utah Mario Capeechi University of Utah See next page for additional authors Follow this and additional works at: https://scholarcommons.sc.edu/ beaufort_naturalsciences_facpub Part of the Biology Commons, and the Genetics Commons Publication Info Published in Development, Volume 141, Issue 19, 2014, pages 3697-3708. © 2014 by the authors. Published by The ompC any of Biologists Ltd. Chojnowski, J. L., Masuda, K., Trau, H. A., Thomas, K., Capecchi, M., & Manley, N. R. (2014). Multiple roles for HOXA3 in regulating thymus and parathyroid differentiation and morphogenesis in mouse. Development (Cambridge, England), 141(19), 3697–3708. http://doi.org/10.1242/dev.110833 This Article is brought to you by the Department of Natural Sciences at Scholar Commons. It has been accepted for inclusion in Faculty Publications by an authorized administrator of Scholar Commons. For more information, please contact [email protected]. Author(s) Jena L. Chojnowski, Kyoko Masuda, Heidi A. Trau, Kirk Thomas, Mario Capeechi, and Nancy R. Manley This article is available at Scholar Commons: https://scholarcommons.sc.edu/beaufort_naturalsciences_facpub/13 Development. 2014 Oct; 141(19): 3697–3708. PMCID: PMC4197593 doi: 10.1242/dev.110833 Multiple roles for HOXA3 in regulating thymus and parathyroid differentiation and morphogenesis in mouse Jena L. Chojnowski,1 Kyoko Masuda,1,* Heidi A.
    [Show full text]