DOCSLIB.ORG

MDGA2 Is a Novel Tumour Suppressor Cooperating with DMAP1 in Gastric

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
MDGA2 Is a Novel Tumour Suppressor Cooperating with DMAP1 in Gastric Gut Online First, published on July 23, 2015 as 10.1136/gutjnl-2015-309276 GI cancer ORIGINAL ARTICLE MDGA2 is a novel tumour suppressor cooperating Gut: first published as 10.1136/gutjnl-2015-309276 on 23 July 2015. Downloaded from with DMAP1 in gastric cancer and is associated with disease outcome Kunning Wang,1 Qiaoyi Liang,1 Xiaoxing Li,1,2 Ho Tsoi,1 Jingwan Zhang,1 Hua Wang,3 Minnie Y Y Go,1 Philip W Y Chiu,4 Enders K W Ng,4 Joseph J Y Sung,1 Jun Yu1 ▸ Additional material is ABSTRACT published online only. To view Background Using the promoter methylation assay, Significance of this study please visit the journal online (http://dx.doi.org/10.1136/ we have shown that MDGA2 (MAM domain containing gutjnl-2015-309276). glycosylphosphatidylinositol anchor 2) is preferentially fi methylated in gastric cancer. We analysed its biological What is already known on this subject? For numbered af liations see fi ▸ end of article. effects and prognostic signi cance in gastric cancer. MDGA2 belongs to the brain-derived Methods MDGA2 methylation status was evaluated by immunoglobulin superfamily and has been Correspondence to combined bisulfite restriction analysis and bisulfite reported to regulate the growth of axons and Professor Jun Yu, Department genomic sequencing. The effects of MDGA2 the development of inhibitory synapse. of Medicine and Therapeutics, re-expression or knockdown on cell proliferation, ▸ State Key Laboratory of Expression of human MDGA2 is detected in Digestive Disease, Institute of apoptosis and the cell cycle were determined. MDGA2 normal human stomach. interacting protein was identified by mass spectrometry Digestive Disease, Prince of fi Wales Hospital, The Chinese and MDGA2-related cancer pathways by reporter activity What are the new ndings? ▸ MDGA2 University of Hong Kong, and PCR array analyses. The clinical impact of MDGA2 is frequently silenced or downregulated Shatin, NT, Hong Kong Hong was assessed in 218 patients with gastric cancer. in gastric cancer by promoter hypermethylation. Kong; [email protected] ▸ MDGA2 Results MDGA2 was commonly silenced in gastric acts as a novel tumour suppressor in KW and QL are joint first cancer cells (10/11) and primary gastric cancers due to gastric cancer through inhibiting cell – authors. promoter hypermethylation. MDGA2 significantly proliferation, suppressing G1 S cell cycle inhibited cell proliferation by causing G1–S cell cycle transition and inducing cell apoptosis. Received 29 January 2015 ▸ The tumour suppressive effect of MDGA2 is http://gut.bmj.com/ Revised 24 June 2015 arrest and inducing cell apoptosis in vitro, and Accepted 27 June 2015 suppressed xenograft tumour growth in both mediated by directly cooperating with DMAP1 subcutaneous and orthotopic xenograft mouse models and consequently inducing p53/p21 signalling (both p<0.001). The anti-tumorigenic effect of MDGA2 cascade. was mediated through direct stabilising of DNA ▸ MDGA2 methylation is an independent methyltransferase 1 associated protein 1 (DMAP1), prognostic biomarker in patients with gastric which played a tumour suppressive role in gastric cancer. cancer. on September 30, 2021 by guest. Protected copyright. This interaction activated their downstream key elements How might it impact on clinical practice in of p53/p21 signalling cascades. Moreover, promoter the foreseeable future? methylation of MDGA2 was detected in 62.4% (136/ ▸ Detection of methylated MDGA2 may serve as 218) of gastric cancers. Multivariate analysis showed a new biomarker for the prognosis of patients that patients with MDGA2 hypermethylation had a with gastric cancer. significantly decreased survival (p=0.005). Kaplan–Meier survival curves showed that MDGA2 hypermethylation was significantly associated with shortened survival in suppressor genes by promoter hypermethylation con- patients with early gastric cancer. 2 Conclusions MDGA2 is a critical tumour suppressor in tributes to the development of gastric cancer. fi gastric carcinogenesis; its hypermethylation is an Identi cation of novel tumour suppressive genes tar- independent prognostic factor in patients with gastric geted by promoter hypermethylation in gastric cancer. cancer may provide insights into the epigenetic mechanisms for the inactivation of tumour suppres- sive pathways. This is also important for the identifi- cation of new markers and therapeutic targets for diagnosis and treatment of this disease. In a previous INTRODUCTION study using MeDIP-chip for a genome-wide screen Gastric cancer is the fifth leading cause of cancer and for hypermethylated candidates in gastric cancer, we To cite: Wang K, Liang Q, the third leading cause of cancer-related mortality found the gene MDGA2 (MAM domain containing Li X, et al. Gut Published Online First: [please include globally. The prognosis of patients with gastric cancer glycosylphosphatidylinositol anchor 2), the function Day Month Year] continues to be poor, despite improving surgical and of which remains largely uncharacterised, was regu- doi:10.1136/gutjnl-2015- adjuvant treatment approaches, with a 5-year overall lated by promoter methylation in gastric cancer 3 309276 survival of less than 25%.1 Inactivation of tumour cells. Wang K, et al. Gut 2015;0:1–13. doi:10.1136/gutjnl-2015-309276 1 Copyright Article author (or their employer) 2015. Produced by BMJ Publishing Group Ltd (& BSG) under licence. GI cancer MDGA2, also known as MAMDC1, is located on chromo- online supplementary table S1). The immune complexes some 14q21.3 and belongs to the brain-derived immunoglobulin were precipitated by centrifugation and separated by SDS-PAGE. superfamily (MDGA1 and MDGA2) in rat.4 MDGAs are classi- Candidate targets were excised from gels and subjected to protein Gut: first published as 10.1136/gutjnl-2015-309276 on 23 July 2015. Downloaded from fied as cell adhesion molecules and have been reported to regu- identification by undertaking the in-gel digestion approach and late the growth of axons and the development of inhibitory using matrix-assisted laser desorption/ionization time-of-flight/ synapses.56However, the function of MDGAs has not yet been time-of-flight (MALDI-TOF/TOF) mass spectrometry.8 investigated in other organs or diseases. Data in the Human Whole cell lysate (150 μg protein) and co-immunoprecipitation Protein Altas show high or medium protein expression of (Co-IP) precipitant by anti-His-tag, anti-DNA methyltransferase 1 MDGA2 in some normal human tissues including the stomach associated protein 1 (DMAP1) antibody or IgG were immuno- (http://www.proteinatlas.org/ENSG00000139915/tissue), but it blotted with either anti-DMAP1 or anti-MDGA2 antibody to was not detected in all 12 tested stomach cancer tissues accord- confirm the interaction of MDGA2 and DMAP1 that was identi- ing to data in the Human Protein Altas (http://www.proteinatlas. fied by mass spectrometry. The lysate (6% input, 10 μgprotein) org/ENSG00000139915/cancer). These findings collectively wasalsousedasacontrol. suggest that inactivation of MDGA2 may play a role during gastric carcinogenesis. Glutathione S-transferase protein pull-down assay We conducted the first study on MDGA2 in gastric cancer Purified recombinant human glutathione S-transferase (GST)- with the aim of elucidating the functional significance and DMAP1 protein (Abnova, Taipei City, Taiwan) and His-tagged molecular mechanism of MDGA2, as well as the clinical impli- recombinant human MDGA2 (MDGA2-His; Creative Biomart, cations of its promoter methylation in this malignancy. Shirley, New York, USA) were used for pull-down assay. GST protein was expressed in E coli strain BL21 (DE3), and the GST MATERIALS AND METHODS protein was purified using a GSTrap column (GE Healthcare, Subjects and sample collection Buckinghamshire, UK). GST-DMAP1 and GST proteins (0.2 μg Two hundred and eighteen primary gastric cancer samples each) were captured by 30 μL glutathione sepharose beads (GE were collected during surgical resection at the First Affiliated Healthcare) at 4°C for 2 h; 0.2 μg of MDGA2-His was then Hospital of Sun Yat-sen University in Guangzhou, China. In incubated with the beads loaded with GST-DMAP1 or GST for addition, 22 paired biopsy specimens of gastric tumour and 2 h at 4°C. The beads with protein complexes were then washed adjacent non-tumour sites from patients with gastric cancer and separated by SDS-PAGE and visualised by Coomassie and 20 biopsy specimens of normal gastric mucosa from Brilliant Blue staining. healthy controls were obtained during endoscopy at the Prince of Wales Hospital, The Chinese University of Hong Kong, Ubiquitination assay Hong Kong. None of these patients received preoperative AGS and BGC823 cells stably transfected with MDGA2 expres- chemotherapy or radiotherapy. The samples were all confirmed sion vector or empty vector were incubated in the presence or histologically and all subjects provided informed consent for absence of 30 μM MG132 (Cell Signaling Technology, Danvers, obtaining the study specimens. Human normal stomach cDNA Massachusetts, USA) for 24 h. Total proteins were extracted http://gut.bmj.com/ was purchased commercially (Stratagene, La Jolla, California, using RIPA buffer supplemented with proteinase inhibitor. USA). Immunoprecipitation was then performed using anti-DMAP1 or anti-IgG, respectively. The immunoprecipitated proteins were sub- In vivo subcutaneous and orthotopic xenograft models jected to western blotting using anti-ubiquitin to evaluate the ubi- BGC823 cells (1×107 cells in 0.1 mL phosphate-buffered quitination level. The inputs
Load more

Recommended publications
  • DNA Damage Triggers Tubular Endoplasmic Reticulum Extension to Promote Apoptosis by Facilitating ER-Mitochondria Signaling
    www.nature.com/cr www.cell-research.com ARTICLE OPEN DNA damage triggers tubular endoplasmic reticulum extension to promote apoptosis by facilitating ER-mitochondria signaling Pengli Zheng1,2, Qingzhou Chen1, Xiaoyu Tian1, Nannan Qian1,3, Peiyuan Chai1, Bing Liu1,4, Junjie Hu 5,6, Craig Blackstone2, Desheng Zhu7, Junlin Teng 1 and Jianguo Chen1,4 The endoplasmic reticulum (ER) is composed of the nuclear envelope, perinuclear sheets and a peripheral tubular network. The peripheral ER and mitochondria form tight contacts at specific subdomains, which coordinate the functions of the two organelles and are required for multiple cellular processes such as Ca2+ transfer and apoptosis. However, it is largely unknown how ER morphology and ER-mitochondria signaling are dynamically regulated under different physiological or pathological conditions such as DNA damage. Here we show that the peripheral, tubular ER undergoes significant extension in response to DNA damage, and that this process is dependent on p53-mediated transcriptional activation of the ER-shaping proteins REEP1, REEP2 and EI24 (alias PIG8). This promotes the formation of ER-mitochondria contacts through EI24 and the mitochondrial outer membrane protein VDAC2, facilitates Ca2+ transfer from ER to mitochondria and promotes DNA damage-induced apoptosis. Thus, we identify a unique DNA damage response pathway involving alterations in ER morphology, ER-mitochondria signaling, and apoptosis. Cell Research (2018) 28:833–854; https://doi.org/10.1038/s41422-018-0065-z INTRODUCTION transfer with high efficiency from the ER to mitochondria, which The endoplasmic reticulum (ER) is the largest membranous is necessary for mitochondrial metabolism.16 However, dramati- organelle and performs essential roles in protein synthesis and cally increased ER-mitochondria Ca2+ flux triggers apoptosis by secretion, Ca2+ homeostasis, and lipid metabolism.
    [Show full text]
  • Ei24) Reduced Cell Proliferation and Aggregate-Size in Dictyostelium Discoideum NEHA GUPTA and SHWETA SARAN*
    Int. J. Dev. Biol. 62: 273-283 (2018) https://doi.org/10.1387/ijdb.170327ss www.intjdevbiol.com Deletion of etoposide-induced 2.4 kb transcript (ei24) reduced cell proliferation and aggregate-size in Dictyostelium discoideum NEHA GUPTA and SHWETA SARAN* School of Life Sciences, Jawaharlal Nehru University, New Delhi, India ABSTRACT The etoposide-induced 2.4 kb transcript (ei24) gene is induced both by p53 and etoposide, an anti-cancer tumour drug. There is no p53 gene present in Dictyostelium discoideum. Thus, the functions of ei24 in the absence of p53 were analysed. Both overexpressor (ei24OE) and knockout (ei24-) mutants were made to study its role during growth, development and differentiation. Addi- tionally, cell cycle and its response to DNA-damage were also analysed. We identified, characterized and elucidated the functions of the ei24 gene in Dictyostelium. In silico analyses demonstrated the conservation across eukaryotes and in situ hybridization showed it to be prestalk-specific.ei24 - cells showed reduced cell proliferation and cell-cohesive properties, ultimately forming small-sized ag- gregates that developed into miniature and stalky fruiting bodies. The ei24OE cells formed fruiting bodies with engorged or double-decker type sori with short stalks. The ei24- cells showed reduced cAMP signalling with lower intracellular cAMP levels resulting in diminished migration of cells along cAMP gradients. Deletion of ei24 resulted in mis-expression of prestalk-specific markers. Cell cycle analysis revealed an increased bias towards the stalk-pathway by ei24- cells and vice-versa for ei24OE cells. EI24 in Dictyostelium functions even in the absence of p53 and is induced in response to both UV-radiation and etoposide treatments.
    [Show full text]
  • Experimental Glaucoma and Optic Nerve Transection Induce Simultaneous Upregulation of Proapoptotic and Prosurvival Genes
    Experimental Glaucoma and Optic Nerve Transection Induce Simultaneous Upregulation of Proapoptotic and Prosurvival Genes Hani Levkovitch-Verbin, Rima Dardik, Shelly Vander, Yael Nisgav, Maya Kalev-Landoy, and Shlomo Melamed PURPOSE. To investigate changes in gene expression induced by imental glaucoma allow identification of genes and proteins elevated intraocular pressure (IOP) and complete optic nerve that are involved in different stages of the disease.3–7 Extensive transsection (ONT) over time. data have been collected to understand how RGCs degenerate METHODS. A gene array of 18 signal transduction pathways was in glaucoma. Gene array analysis recently revealed up- and downregulation of many genes in response to elevated intraoc- used to examine the changes in RNA profiles of retinas post- 8–10 10 ONT in rats. Among the seven genes that were determined to ular pressure (IOP). Ahmed et al. analyzed early (8 days) be upregulated, four were confirmed to have higher expres- and late (35 days) changes in gene expression of retinas from sion by semiquantitative RT-PCR analysis: Ei24 and Gadd45a glaucomatous eyes of rats, by using the episcleral vein injection (both associated with apoptosis induced via the p53 pathway), of hypertonic saline. They found altered expression of 81 genes, approximately half of which were associated with ei- IAP-1 (inhibitor of apoptosis protein 1), and Cdk2 (cell cycle 8 regulation and apoptosis). Their mRNA levels were then stud- ther apoptosis or neuroinflammatory responses. Farkas et al. ied by quantitative RT-PCR in experimental glaucoma and ONT found increased mRNA levels of iron-regulating proteins, sug- over time. Levels of the corresponding proteins were evaluated gesting the involvement of iron and copper metabolism and by Western blot analysis and immunohistochemistry.
    [Show full text]
  • Retinoic Acid Receptor-Independent Mechanism of Apoptosis of Melanoma Cells by the Retinoid CD437 (AHPN)
    Cell Death and Differentiation (2001) 8, 878 ± 886 ã 2001 Nature Publishing Group All rights reserved 1350-9047/01 $15.00 www.nature.com/cdd Retinoic acid receptor-independent mechanism of apoptosis of melanoma cells by the retinoid CD437 (AHPN) 1 1 1 2 3 X Zhao , K Demary , L Wong , C Vaziri , AB McKenzie , Introduction TJ Eberlein4 and RA Spanjaard*,1 Retinoic acid (RA)3 and other retinoids are small molecules 1 Department of Otolaryngology, Cancer Research Center, Boston University derived from retinol (Vitamin A) which guide proper School of Medicine, Boston, MA, USA mammalian embryonic development by activating RARs, a 2 Department of Medicine, Cancer Research Center, Boston University School of class of ligand-dependent transcription factors which regulate Medicine, Boston, MA, USA gene expression through binding to RA response elements 3 Department of Surgery, Brigham and Women's Hospital, Harvard Medical (RAREs).1,2 RA can also induce growth-arrest and differentia- School, Boston, MA, USA 3 4 Department of Surgery, Washington University School of Medicine, St. Louis, tion of a large variety of tumor cells in vitro and mediate 4 MO, USA similar effects in some neoplastic tissues, which is the * Corresponding author: Boston University School of Medicine, Cancer rationale behind the use of retinoids as pharmacological Research Center, 715 Albany Street R903, Boston, MA 02118. agents for differentiation and chemoprevention therapies in Tel: (617) 638-4811; Fax: (617) 638-5837; E-mail: [email protected] certain cancers.5,6 In order to understand the molecular basis of the effects Received 18.1.01; revised 20.3.01; accepted 9.4.01 of retinoids on the genetic program of tumor cells, we have Edited by D Green utilized the S91 murine melanoma cell line as a model system for the etiology of this disease.
    [Show full text]
  • 42328 EI24 (D3F6Z) Rabbit Mab
    Revision 1 C 0 2 - t EI24 (D3F6Z) Rabbit mAb a e r o t S Orders: 877-616-CELL (2355) [email protected] 8 Support: 877-678-TECH (8324) 2 3 Web: [email protected] 2 www.cellsignal.com 4 # 3 Trask Lane Danvers Massachusetts 01923 USA For Research Use Only. Not For Use In Diagnostic Procedures. Applications: Reactivity: Sensitivity: MW (kDa): Source/Isotype: UniProt ID: Entrez-Gene Id: WB, IP H M R Endogenous 30 Rabbit IgG O14681 9538 Product Usage Information 8. Zhao, Y.G. et al. (2012) J Biol Chem 287, 42053-63. 9. Devkota, S. et al. (2016) Autophagy 12, 2038-2053. Application Dilution 10. Choi, J.M. et al. (2013) Oncotarget 4, 2383-96. Western Blotting 1:1000 Immunoprecipitation 1:200 Storage Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody. Specificity / Sensitivity EI24 (D3F6Z) Rabbit mAb recognizes endogenous levels of total EI24 protein. Species Reactivity: Human, Mouse, Rat Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala31 of human EI24 protein. Background Etoposide-induced 2.4 mRNA (EI24)/p53-induced gene 8 (PIG8) was identified as a DNA damage response gene induced by etoposide in a p53 dependent manner with roles in growth suppression and apoptosis (1-3). As a pro-apoptotic gene, some evidence suggests that EI24 functions as a tumor suppressor gene in cases such as breast and cervical cancer (4-6).
    [Show full text]
  • Potential Whole-Cell Biosensors for Detection of Metal Using Merr Family Proteins from Enterobacter Sp
    micromachines Article Potential Whole-Cell Biosensors for Detection of Metal Using MerR Family Proteins from Enterobacter sp. YSU and Stenotrophomonas maltophilia OR02 Georgina Baya 1, Stephen Muhindi 2, Valentine Ngendahimana 3 and Jonathan Caguiat 1,* 1 Department of Biological and Chemical Sciences, Youngstown State University, Youngstown, OH 44555, USA; [email protected] 2 Department of Biological Sciences, University of Toledo, Toledo, OH 43606, USA; [email protected] 3 Biology Department, Lone Star College-CyFair, 9191 Barker Cypress Rd, Cypress, TX 77433, USA; [email protected] * Correspondence: [email protected]; Tel.: +1-330-941-2063 Abstract: Cell-based biosensors harness a cell’s ability to respond to the environment by repurposing its sensing mechanisms. MerR family proteins are activator/repressor switches that regulate the expression of bacterial metal resistance genes and have been used in metal biosensors. Upon metal binding, a conformational change switches gene expression from off to on. The genomes of the multi- metal resistant bacterial strains, Stenotrophomonas maltophilia Oak Ridge strain 02 (S. maltophilia 02) and Enterobacter sp. YSU, were recently sequenced. Sequence analysis and gene cloning identified three mercury resistance operons and three MerR switches in these strains. Transposon mutagenesis and sequence analysis identified Enterobacter sp. YSU zinc and copper resistance operons, which ap- pear to be regulated by the protein switches, ZntR and CueR, respectively. Sequence analysis and Citation: Baya, G.; Muhindi, S.; reverse transcriptase polymerase chain reaction (RT-PCR) showed that a CueR switch appears to Ngendahimana, V.; Caguiat, J. activate a S. maltophilia 02 copper transport gene in the presence of CuSO4 and HAuCl4·3H2O.
    [Show full text]
  • Mdr1b Facilitates P53-Mediated Cell Death and P53 Is Required for Mdr1b Upregulation in Vivo
    Oncogene (2001) 20, 303 ± 313 ã 2001 Nature Publishing Group All rights reserved 0950 ± 9232/01 $15.00 www.nature.com/onc Mdr1b facilitates p53-mediated cell death and p53 is required for Mdr1b upregulation in vivo Valerie Lecureur1, Jaideep V Thottassery1, Daxi Sun1, Erin G Schuetz1, Jill Lahti2, Gerard P Zambetti3 and John D Schuetz*,1 1Department of Pharmaceutical Sciences, St Jude Children's Research Hospital, 332 North Lauderdale Avenue, Memphis, Tennessee, TN 38105, USA; 2Department of Tumor Cell Biology, St Jude Children's Research Hospital, 332 North Lauderdale Avenue, Memphis, Tennessee, TN 38105, USA; 3Department of Biochemistry, St Jude Children's Research Hospital, 332 North Lauderdale Avenue, Memphis, Tennessee, TN 38105, USA The mdr1b gene is thought to be a ``stress-responsive'' share greater than 88% identity, are often expressed in gene, however it is unknown if this gene is regulated by the same tissues and can be selectively overproduced p53 in the whole animal. Moreover, it is unknown if during selection for resistance to cytotoxic agents (e.g., overexpression of mdr1b aects cell survival. The taxol, vinblastine, adriamycin (Lothstein et al., 1989)). dependence of mdr1b upon p53 for upregulation was Further, selection for drug resistance with ectopically evaluated in p53 knockout mice. Wild-type (wt) or introduced mdr1a or mdr1b expression vectors yields p537/7 mice were treated singly or in combination similar drug resistance pro®les for many drugs with gamma irradiation (IR) and/or the potent DNA (Devault and Gros, 1990). In contrast, the application damaging agent, diethylnitrosoamine (DEN). Both IR of cytotoxic drug selective pressure to murine macro- and DEN induced mdr1b in wild-type animals, but not in phage cells caused a switch in endogenous mdr the p537/7 mice.
    [Show full text]
  • Genenames.Org: the HGNC Resources in 2011 Ruth L
    Nucleic Acids Research Advance Access published October 6, 2010 Nucleic Acids Research, 2010, 1–6 doi:10.1093/nar/gkq892 genenames.org: the HGNC resources in 2011 Ruth L. Seal*, Susan M. Gordon, Michael J. Lush, Mathew W. Wright and Elspeth A. Bruford European Bioinformatics Institute (EMBL-EBI), Wellcome Trust Genome Campus, Hinxton, Cambridgeshire, CB10 1SA, UK Received September 15, 2010; Accepted September 21, 2010 ABSTRACT In order to achieve this, we endeavour to contact the The HUGO Gene Nomenclature Committee (HGNC) researchers that work on particular genes for their Downloaded from aims to assign a unique gene symbol and name to advice and input before approving symbols, and encour- every human gene. The HGNC database currently age researchers to submit proposed gene symbols directly contains almost 30 000 approved gene symbols, to us to determine their suitability prior to publication. over 19 000 of which represent protein-coding The HGNC team attends conferences regularly to ensure that we are meeting the requirements of the community genes. The public website, www.genenames.org, and to discuss the nomenclature of specific gene families nar.oxfordjournals.org displays all approved nomenclature within Symbol and locus types. We work closely with the nomenclature Reports that contain data curated by HGNC editors committees for several other species, especially the mouse and links to related genomic, phenotypic and prote- (2), rat (3), zebrafish (4) and Xenopus (5) to ensure that omic information. Here we describe improvements orthologous vertebrate genes are assigned equivalent to our resources, including a new Quick Gene symbols wherever possible. HGNC symbols are used by Search, a new List Search, an integrated HGNC most biomedical databases, including Ensembl (6), Vega BioMart and a new Statistics and Downloads facility.
    [Show full text]
  • EI24 (NM 004879) Human Untagged Clone – SC117074 | Origene
    OriGene Technologies, Inc. 9620 Medical Center Drive, Ste 200 Rockville, MD 20850, US Phone: +1-888-267-4436 [email protected] EU: [email protected] CN: [email protected] Product datasheet for SC117074 EI24 (NM_004879) Human Untagged Clone Product data: Product Type: Expression Plasmids Product Name: EI24 (NM_004879) Human Untagged Clone Tag: Tag Free Symbol: EI24 Synonyms: EPG4; PIG8; TP53I8 Vector: pCMV6-XL5 E. coli Selection: Ampicillin (100 ug/mL) Cell Selection: None Fully Sequenced ORF: >OriGene ORF within SC117074 sequence for NM_004879 edited (data generated by NextGen Sequencing) ATGGCTGACAGTGTCAAAACCTTTCTCCAGGACCTTGCCAGAGGAATCAAAGACTCCATC TGGGGTATTTGTACCATCTCAAAGCTAGATGCTCGAATCCAGCAAAAGAGAGAGGAGCAG CGTCGAAGAAGGGCAAGTAGTGTCTTGGCACAGAGAAGAGCCCAGAGTATAGAGCGGAAG CAAGAGAGTGAGCCACGTATTGTTAGTAGAATTTTCCAGTGTTGTGCTTGGAATGGTGGA GTGTTCTGGTTCAGTCTCCTCTTGTTTTATCGAGTATTTATTCCTGTGCTTCAGTCGGTA ACAGCCCGAATTATCGGTGACCCATCACTACATGGAGATGTTTGGTCGTGGCTGGAATTC TTCCTCACGTCAATTTTCAGTGCTCTTTGGGTGCTCCCCTTGTTTGTGCTTAGCAAAGTG GTGAATGCCATTTGGTTTCAGGATATAGCTGACCTGGCATTTGAGGTATCAGGGAGGAAG CCTCACCCATTCCCTAGTGTCAGCAAAATAATTGCTGACATGCTCTTCAACCTTTTGCTG CAGGCTCTTTTCCTCATTCAGGGAATGTTTGTGAGTCTCTTTCCCATCCATCTTGTCGGT CAGCTGGTTAGTCTCCTGCATATGTCCCTTCTCTACTCACTGTACTGCTTTGAATATCGT TGGTTCAATAAAGGAATTGAAATGCACCAGCGGTTGTCTAACATAGAAAGGAATTGGCCT TACTACTTTGGGTTTGGTTTGCCCTTGGCTTTTCTCACAGCAATGCAGTCCTCATATATT ATCAGTGGCTGCCTTTTCTCTATCCTCTTTCCTTTATTCATTATCAGCGCCAATGAAGCA AAGACCCCTGGCAAAGCGTATCTCTTCCAGTTGCGCCTCTTCTCCTTGGTGGTCTTCTTA AGCAACAGACTCTTCCACAAGACAGTCTACCTGCAGTCGGCCCTGAGCAGCTCTACTTCT
    [Show full text]
  • New Approaches for Quantitative Reconstruction of Radiation Dose in Human Blood Cells Shanaz A
    www.nature.com/scientificreports OPEN New Approaches for Quantitative Reconstruction of Radiation Dose in Human Blood Cells Shanaz A. Ghandhi 1,2*, Igor Shuryak1,2, Shad R. Morton1, Sally A. Amundson 1 & David J. Brenner1 In the event of a nuclear attack or large-scale radiation event, there would be an urgent need for assessing the dose to which hundreds or thousands of individuals were exposed. Biodosimetry approaches are being developed to address this need, including transcriptomics. Studies have identifed many genes with potential for biodosimetry, but, to date most have focused on classifcation of samples by exposure levels, rather than dose reconstruction. We report here a proof-of-principle study applying new methods to select radiation-responsive genes to generate quantitative, rather than categorical, radiation dose reconstructions based on a blood sample. We used a new normalization method to reduce efects of variability of signal intensity in unirradiated samples across studies; developed a quantitative dose-reconstruction method that is generally under-utilized compared to categorical methods; and combined these to determine a gene set as a reconstructor. Our dose-reconstruction biomarker was trained using two data sets and tested on two independent ones. It was able to reconstruct dose up to 4.5 Gy with root mean squared error (RMSE) of ± 0.35 Gy on a test dataset using the same platform, and up to 6.0 Gy with RMSE of ± 1.74 Gy on a test set using a diferent platform. In the event of a nuclear attack or large-scale radiation event, there would be an urgent need for assessing the dose to which hundreds or thousands of individuals were exposed1–4.
    [Show full text]
  • Gene-Expression and in Vitro Function of Mesenchymal Stromal Cells Are Affected in Juvenile Myelomonocytic Leukemia
    Myeloproliferative Disorders SUPPLEMENTARY APPENDIX Gene-expression and in vitro function of mesenchymal stromal cells are affected in juvenile myelomonocytic leukemia Friso G.J. Calkoen, 1 Carly Vervat, 1 Else Eising, 2 Lisanne S. Vijfhuizen, 2 Peter-Bram A.C. ‘t Hoen, 2 Marry M. van den Heuvel-Eibrink, 3,4 R. Maarten Egeler, 1,5 Maarten J.D. van Tol, 1 and Lynne M. Ball 1 1Department of Pediatrics, Immunology, Hematology/Oncology and Hematopoietic Stem Cell Transplantation, Leiden University Med - ical Center, the Netherlands; 2Department of Human Genetics, Leiden University Medical Center, Leiden, the Netherlands; 3Dutch Childhood Oncology Group (DCOG), The Hague, the Netherlands; 4Princess Maxima Center for Pediatric Oncology, Utrecht, the Nether - lands; and 5Department of Hematology/Oncology and Hematopoietic Stem Cell Transplantation, Hospital for Sick Children, University of Toronto, ON, Canada ©2015 Ferrata Storti Foundation. This is an open-access paper. doi:10.3324/haematol.2015.126938 Manuscript received on March 5, 2015. Manuscript accepted on August 17, 2015. Correspondence: [email protected] Supplementary data: Methods for online publication Patients Children referred to our center for HSCT were included in this study according to a protocol approved by the institutional review board (P08.001). Bone-marrow of 9 children with JMML was collected prior to treatment initiation. In addition, bone-marrow after HSCT was collected from 5 of these 9 children. The patients were classified following the criteria described by Loh et al.(1) Bone-marrow samples were sent to the JMML-reference center in Freiburg, Germany for genetic analysis. Bone-marrow samples of healthy pediatric hematopoietic stem cell donors (n=10) were used as control group (HC).
    [Show full text]
  • EI24 Antibody Cat
    EI24 Antibody Cat. No.: 6487 EI24 Antibody Immunohistochemistry of EI24 in rat liver tissue with EI24 antibody at 5 μg/mL. Specifications HOST SPECIES: Rabbit SPECIES REACTIVITY: Human, Mouse, Rat HOMOLOGY: Predicted species reactivity based on immunogen sequence: Bovine: (95%) EI24 antibody was raised against a 19 amino acid synthetic peptide near the carboxy terminus of human EI24. IMMUNOGEN: The immunogen is located within the last 50 amino acids of EI24. TESTED APPLICATIONS: ELISA, IHC-P, WB September 29, 2021 1 https://www.prosci-inc.com/ei24-antibody-6487.html EI24 antibody can be used for detection of EI24 by Western blot at 1 - 2 μg/mL. Antibody can also be used for immunohistochemistry starting at 5 μg/mL. APPLICATIONS: Antibody validated: Western Blot in rat samples and Immunohistochemistry in rat samples. All other applications and species not yet tested. At least two isoforms of EI24 are known to exist; this antibody will only detect the larger SPECIFICITY: isoform. POSITIVE CONTROL: 1) Cat. No. 1464 - Rat Liver Tissue Lysate Properties PURIFICATION: EI24 Antibody is affinity chromatography purified via peptide column. CLONALITY: Polyclonal ISOTYPE: IgG CONJUGATE: Unconjugated PHYSICAL STATE: Liquid BUFFER: EI24 Antibody is supplied in PBS containing 0.02% sodium azide. CONCENTRATION: 1 mg/mL EI24 antibody can be stored at 4˚C for three months and -20˚C, stable for up to one year. STORAGE CONDITIONS: As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures. Additional Info OFFICIAL SYMBOL: EI24 ALTERNATE NAMES: EI24 Antibody: EPG4, PIG8, TP53I8, p53-induced gene 8 protein ACCESSION NO.: NP_004870 PROTEIN GI NO.: 55956766 GENE ID: 9538 USER NOTE: Optimal dilutions for each application to be determined by the researcher.
    [Show full text]