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Microglia Emerge from Erythromyeloid Precursors Via Pu.1- and Irf8-Dependent Pathways
ART ic LE S Microglia emerge from erythromyeloid precursors via Pu.1- and Irf8-dependent pathways Katrin Kierdorf1,2, Daniel Erny1, Tobias Goldmann1, Victor Sander1, Christian Schulz3,4, Elisa Gomez Perdiguero3,4, Peter Wieghofer1,2, Annette Heinrich5, Pia Riemke6, Christoph Hölscher7,8, Dominik N Müller9, Bruno Luckow10, Thomas Brocker11, Katharina Debowski12, Günter Fritz1, Ghislain Opdenakker13, Andreas Diefenbach14, Knut Biber5,15, Mathias Heikenwalder16, Frederic Geissmann3,4, Frank Rosenbauer6 & Marco Prinz1,17 Microglia are crucial for immune responses in the brain. Although their origin from the yolk sac has been recognized for some time, their precise precursors and the transcription program that is used are not known. We found that mouse microglia were derived from primitive c-kit+ erythromyeloid precursors that were detected in the yolk sac as early as 8 d post conception. + lo − + − + These precursors developed into CD45 c-kit CX3CR1 immature (A1) cells and matured into CD45 c-kit CX3CR1 (A2) cells, as evidenced by the downregulation of CD31 and concomitant upregulation of F4/80 and macrophage colony stimulating factor receptor (MCSF-R). Proliferating A2 cells became microglia and invaded the developing brain using specific matrix metalloproteinases. Notably, microgliogenesis was not only dependent on the transcription factor Pu.1 (also known as Sfpi), but also required Irf8, which was vital for the development of the A2 population, whereas Myb, Id2, Batf3 and Klf4 were not required. Our data provide cellular and molecular insights into the origin and development of microglia. Microglia are the tissue macrophages of the brain and scavenge dying have the ability to give rise to microglia and macrophages in vitro cells, pathogens and molecules using pattern recognition receptors and in vivo under defined conditions. -
The Transcription Factors C-Myb and GATA-2 Act Independently in The
Proc. Natl. Acad. Sci. USA Vol. 93, pp. 5313-5318, May 1996 Medical Sciences The transcription factors c-myb and GATA-2 act independently in the regulation of normal hematopoiesis PAOLA MELOTrl AND BRUNO CALABRETTA Department of Microbiology and Immunology, Kimmel Cancer Institute, Thomas Jefferson University, 233 South 10th Street, Philadelphia, PA 19107 Communicated by Sidney Weinhouse, Thomas Jefferson University, Philadelphia, PA, January 23, 1996 (received for review, October 20, 1995) ABSTRACT The transcription factors c-myb and GATA-2 erythromyeloid differentiation (7). This process appears to are both required for blood cell development in vivo and in rest in the ability of c-myb to activate the expression of vitro. However, very little is known on their mechanism(s) of hematopoiesis-specific targets such as c-kit,flt-3, GATA-1, and action and whether they impact on complementary or over- CD34, but not GATA-2 (7). The induction of c-kit and flt-3 lapping pathways of hematopoietic proliferation and differ- expression and the dependence of c-myb-transfected ES cells entiation. We report here that embryonic stem (ES) cells on the expression of these cytokine receptors for their prolif- transfected with c-myb or GATA-2 cDNAs, individually or in eration (7) strongly suggest that the up-regulation of growth combination, underwent hematopoietic commitment and dif- factor receptor levels is of fundamental importance for the ferentiation in the absence of added hematopoietic growth expansion of progenitor cells. In turn, such a process is factors but that stimulation with c-kit and flt-3 ligands en- probably a requirement for completion of the differentiation hanced colony formation only in the c-myb transfectants. -
Cells Transformed by a V-Myb-Estrogen Receptor Fusion Differentiate Into Multinucleated Giant Cells
JOURNAL OF VIROLOGY, May 1997, p. 3760–3766 Vol. 71, No. 5 0022-538X/97/$04.0010 Copyright q 1997, American Society for Microbiology Cells Transformed by a v-Myb-Estrogen Receptor Fusion Differentiate into Multinucleated Giant Cells UTE ENGELKE, DUEN-MEI WANG, AND JOSEPH S. LIPSICK* Department of Pathology, Stanford University School of Medicine, Stanford, California 94305-5324 Received 22 October 1996/Accepted 29 January 1997 In order to make conditional alleles of the v-myb oncogene, we constructed and tested avian retroviruses which produce a number of different fusion proteins between v-Myb and the human estrogen receptor (ER). We found that the portion of the ER used in making these fusions profoundly affected their transcriptional activation. However, all the fusions tested were only weakly transforming in embryonic yolk sac assays and there was no direct correlation between the level of transcriptional activation and strength of oncogenic transformation. Nevertheless, transformation by a v-Myb-ER fusion was estrogen dependent, and upon with- drawal of the hormone, monocytic-lineage cells differentiated into multinucleated giant cells. Surprisingly, the withdrawal of estrogen caused a dramatic increase in the stability of the fusion protein, although it remained unable to promote cell growth or block differentiation. Conditional alleles of retroviral oncogenes have provided genes, have been isolated and analyzed in considerable detail powerful tools with which to dissect the mechanism of onco- (3, 4). A differential cDNA screen has identified a target gene, genic transformation and the biology of transformed cells (17, mim-1, which is directly activated by the Gag-Myb-Ets protein 29). -
Anti-NFYC Antibody (ARG66549)
Product datasheet [email protected] ARG66549 Package: 100 μl anti-NFYC antibody Store at: -20°C Summary Product Description Rabbit Polyclonal antibody recognizes NFYC Tested Reactivity Hu, Ms Tested Application IHC-P, WB Host Rabbit Clonality Polyclonal Isotype IgG Target Name NFYC Antigen Species Human Immunogen KLH-conjugated synthetic peptide encompassing a sequence within the N-term region of Human NFYC. Conjugation Un-conjugated Alternate Names HAP5; NF-YC; Nuclear transcription factor Y subunit gamma; Nuclear transcription factor Y subunit C; CBF-C; HSM; CBFC; Transactivator HSM-1/2; CAAT box DNA-binding protein subunit C; H1TF2A Application Instructions Application table Application Dilution IHC-P 1:100 - 1:200 WB 1:500 - 1:1000 Application Note IHC-P: Antigen Retrieval: Heat mediation was performed in Sodium citrate buffer (pH 6.0). * The dilutions indicate recommended starting dilutions and the optimal dilutions or concentrations should be determined by the scientist. Calculated Mw 50 kDa Observed Size 50 kDa Properties Form Liquid Purification Affinity purification with immunogen. Buffer 0.42% Potassium phosphate (pH 7.3), 0.87% NaCl, 0.01% Sodium azide and 30% Glycerol. Preservative 0.01% Sodium azide Stabilizer 30% Glycerol Storage instruction For continuous use, store undiluted antibody at 2-8°C for up to a week. For long-term storage, aliquot and store at -20°C. Storage in frost free freezers is not recommended. Avoid repeated freeze/thaw cycles. Suggest spin the vial prior to opening. The antibody solution should be gently mixed before use. www.arigobio.com 1/2 Note For laboratory research only, not for drug, diagnostic or other use. -
A Flexible Microfluidic System for Single-Cell Transcriptome Profiling
www.nature.com/scientificreports OPEN A fexible microfuidic system for single‑cell transcriptome profling elucidates phased transcriptional regulators of cell cycle Karen Davey1,7, Daniel Wong2,7, Filip Konopacki2, Eugene Kwa1, Tony Ly3, Heike Fiegler2 & Christopher R. Sibley 1,4,5,6* Single cell transcriptome profling has emerged as a breakthrough technology for the high‑resolution understanding of complex cellular systems. Here we report a fexible, cost‑efective and user‑ friendly droplet‑based microfuidics system, called the Nadia Instrument, that can allow 3′ mRNA capture of ~ 50,000 single cells or individual nuclei in a single run. The precise pressure‑based system demonstrates highly reproducible droplet size, low doublet rates and high mRNA capture efciencies that compare favorably in the feld. Moreover, when combined with the Nadia Innovate, the system can be transformed into an adaptable setup that enables use of diferent bufers and barcoded bead confgurations to facilitate diverse applications. Finally, by 3′ mRNA profling asynchronous human and mouse cells at diferent phases of the cell cycle, we demonstrate the system’s ability to readily distinguish distinct cell populations and infer underlying transcriptional regulatory networks. Notably this provided supportive evidence for multiple transcription factors that had little or no known link to the cell cycle (e.g. DRAP1, ZKSCAN1 and CEBPZ). In summary, the Nadia platform represents a promising and fexible technology for future transcriptomic studies, and other related applications, at cell resolution. Single cell transcriptome profling has recently emerged as a breakthrough technology for understanding how cellular heterogeneity contributes to complex biological systems. Indeed, cultured cells, microorganisms, biopsies, blood and other tissues can be rapidly profled for quantifcation of gene expression at cell resolution. -
Genome-Wide DNA Methylation Analysis of KRAS Mutant Cell Lines Ben Yi Tew1,5, Joel K
www.nature.com/scientificreports OPEN Genome-wide DNA methylation analysis of KRAS mutant cell lines Ben Yi Tew1,5, Joel K. Durand2,5, Kirsten L. Bryant2, Tikvah K. Hayes2, Sen Peng3, Nhan L. Tran4, Gerald C. Gooden1, David N. Buckley1, Channing J. Der2, Albert S. Baldwin2 ✉ & Bodour Salhia1 ✉ Oncogenic RAS mutations are associated with DNA methylation changes that alter gene expression to drive cancer. Recent studies suggest that DNA methylation changes may be stochastic in nature, while other groups propose distinct signaling pathways responsible for aberrant methylation. Better understanding of DNA methylation events associated with oncogenic KRAS expression could enhance therapeutic approaches. Here we analyzed the basal CpG methylation of 11 KRAS-mutant and dependent pancreatic cancer cell lines and observed strikingly similar methylation patterns. KRAS knockdown resulted in unique methylation changes with limited overlap between each cell line. In KRAS-mutant Pa16C pancreatic cancer cells, while KRAS knockdown resulted in over 8,000 diferentially methylated (DM) CpGs, treatment with the ERK1/2-selective inhibitor SCH772984 showed less than 40 DM CpGs, suggesting that ERK is not a broadly active driver of KRAS-associated DNA methylation. KRAS G12V overexpression in an isogenic lung model reveals >50,600 DM CpGs compared to non-transformed controls. In lung and pancreatic cells, gene ontology analyses of DM promoters show an enrichment for genes involved in diferentiation and development. Taken all together, KRAS-mediated DNA methylation are stochastic and independent of canonical downstream efector signaling. These epigenetically altered genes associated with KRAS expression could represent potential therapeutic targets in KRAS-driven cancer. Activating KRAS mutations can be found in nearly 25 percent of all cancers1. -
The Role of ATP Binding Cassette Transporters in Tissue Defense and Organ Regeneration
0022-3565/09/3281-3–9$20.00 THE JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS Vol. 328, No. 1 Copyright © 2009 by The American Society for Pharmacology and Experimental Therapeutics 132225/3408067 JPET 328:3–9, 2009 Printed in U.S.A. Perspectives in Pharmacology The Role of ATP Binding Cassette Transporters in Tissue Defense and Organ Regeneration Miriam Huls, Frans G. M. Russel, and Rosalinde Masereeuw Department of Pharmacology and Toxicology, Radboud University Nijmegen Medical Centre, Nijmegen Centre for Molecular Life Sciences, The Netherlands Downloaded from Received July 22, 2008; accepted September 11, 2008 ABSTRACT ATP binding cassette (ABC) transporters are ATP-dependent tissues. The expression levels of BCRP and P-gp are tightly con- membrane proteins predominantly expressed in excretory organs, trolled and may determine the differentiation of SP cells toward jpet.aspetjournals.org such as the liver, intestine, blood-brain barrier, blood-testes bar- other more specialized cell types. Although their exact function in rier, placenta, and kidney. Here, they play an important role in these cells is still not clear, they may protect the cells by pumping the absorption, distribution, and excretion of drugs, xenobiotics, out toxicants and harmful products of oxidative stress. Transplan- and endogenous compounds. In addition, the ABC transporters, tation studies in animals revealed that bone marrow-derived SP P-glycoprotein (P-gp/ABCB1) and breast cancer resistance cells contribute to organ repopulation and tissue repair after dam- protein (BCRP/ABCG2), are highly expressed in a population of age, e.g., in liver and heart. The role of SP cells in regeneration of primitive stem cells: the side population (SP). -
Chain Gene Induced by GM-CSF Β Receptor Regulation of Human High-Affinity Ige Molecular Mechanisms for Transcriptional
Molecular Mechanisms for Transcriptional Regulation of Human High-Affinity IgE Receptor β-Chain Gene Induced by GM-CSF This information is current as Kyoko Takahashi, Natsuko Hayashi, Shuichi Kaminogawa of September 23, 2021. and Chisei Ra J Immunol 2006; 177:4605-4611; ; doi: 10.4049/jimmunol.177.7.4605 http://www.jimmunol.org/content/177/7/4605 Downloaded from References This article cites 39 articles, 16 of which you can access for free at: http://www.jimmunol.org/content/177/7/4605.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on September 23, 2021 *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2006 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Molecular Mechanisms for Transcriptional Regulation of Human High-Affinity IgE Receptor -Chain Gene Induced by GM-CSF1 Kyoko Takahashi,*† Natsuko Hayashi,*‡ Shuichi Kaminogawa,† and Chisei Ra2* The -chain of the high-affinity receptor for IgE (FcRI) plays an important role in regulating activation of FcRI-expressing cells such as mast cells in allergic reactions. -
Hematopoietic Stem Cell Marker Antibody Panel (ABCG2, CD34, Store At: Prominin1)
Product datasheet [email protected] ARG30135 Package: 1 pair Hematopoietic Stem Cell Marker Antibody Panel (ABCG2, CD34, Store at: Prominin1) Component Cat No Component Name Host clonality Reactivity Application Package ARG62820 anti-CD34 antibody Mouse mAb Hu Blocking, FACS, 50 μg [4H11(APG)] ICC/IF ARG54138 anti-Prominin-1 Mouse mAb Hu WB 50 μl antibody [6H10-F1-C11] ARG51346 anti-ABCG2 / CD338 Rabbit pAb Hu WB 50 μl antibody ARG65350 Goat anti-Mouse IgG Goat pAb ELISA, IHC, WB 50 μl antibody (HRP) ARG65351 Goat anti-Rabbit IgG Goat pAb Rb ELISA, IHC, WB 50 μl antibody (HRP) Summary Product Description Hematopoietic Stem Cells (HSCs) are the mesodermal progenitor cells that give rise to all types of blood cells including monocytes, macrophages, neutrophils, basophils, eosinophils, erythrocytes and other cells related to lymphoid lineage. It has been a challenge for researchers using HSCs because of the difficulties to isolate them from a large pool of cells because the number of HSCs in each organism is scarce and they appear very similar to lymphocytes morphologically. CD34, CD133 and ABCG2 are reliable marker for HSCs. Using the antibodies included in arigo’s Hematopoietic Stem Cell Panel, researchers can easily identify HSCs from the blood cell population. Sutherland, D.R. et al. (1992) J. Hematother.1:115. Yin, A.H. et al. (1997) Blood 90:5002. Gehling, U.M. et al. (2000) Blood 95:3106. Zhou, S. et al. (2001) Nat. Med. 7:1028. Kim, M. et al. (2002) Clin. Cancer Res. 8:22. www.arigobio.com 1/2 Images ARG51346 anti-ABCG2 / CD338 antibody WB validated image Western Blot: extract from HL-60 cells stained with anti-ABCG2 / CD338 antibody ARG51346 ARG54138 anti-Prominin-1 antibody [6H10-F1-C11] WB validated image Western Blot: Prominin-1 in CaCo2 cell lysate stained with Prominin-1 antibody [6H10-F1-C11] (ARG54138) (diluted in 1: 1000). -
The Id-Protein Family in Developmental and Cancer-Associated Pathways Cornelia Roschger and Chiara Cabrele*
Roschger and Cabrele Cell Communication and Signaling (2017) 15:7 DOI 10.1186/s12964-016-0161-y REVIEW Open Access The Id-protein family in developmental and cancer-associated pathways Cornelia Roschger and Chiara Cabrele* Abstract Inhibitors of DNA binding and cell differentiation (Id) proteins are members of the large family of the helix-loop- helix (HLH) transcription factors, but they lack any DNA-binding motif. During development, the Id proteins play a key role in the regulation of cell-cycle progression and cell differentiation by modulating different cell-cycle regulators both by direct and indirect mechanisms. Several Id-protein interacting partners have been identified thus far, which belong to structurally and functionally unrelated families, including, among others, the class I and II bHLH transcription factors, the retinoblastoma protein and related pocket proteins, the paired-box transcription factors, and the S5a subunit of the 26 S proteasome. Although the HLH domain of the Id proteins is involved in most of their protein-protein interaction events, additional motifs located in their N-terminal and C-terminal regions are required for the recognition of diverse protein partners. The ability of the Id proteins to interact with structurally different proteins is likely to arise from their conformational flexibility: indeed, these proteins contain intrinsically disordered regions that, in the case of the HLH region, undergo folding upon self- or heteroassociation. Besides their crucial role for cell-fate determination and cell-cycle progression during development, other important cellular events have been related to the Id-protein expression in a number of pathologies. Dysregulated Id-protein expression has been associated with tumor growth, vascularization, invasiveness, metastasis, chemoresistance and stemness, as well as with various developmental defects and diseases. -
Transcription Regulation of MYB: a Potential and Novel Therapeutic Target in Cancer
Review Article Page 1 of 11 Transcription regulation of MYB: a potential and novel therapeutic target in cancer Partha Mitra1,2 1Pre-clinical Division, Vaxxas Pty. Ltd. Translational Research Institute, Woolloongabba QLD 4102, Australia; 2Institute of Health and Biomedical Innovation, Queensland University of Technology, Translational Research Institute, Woolloongabba QLD 4102, Australia Correspondence to: Partha Mitra. Pre-clinical Division, Vaxxas Pty. Ltd. Translational Research Institute, 37 Kent St., Woolloongabba QLD 4102, Australia; Queensland University of Technology, Translational Research Institute, 37 Kent St., Woolloongabba QLD 4102, Australia. Email: [email protected]; [email protected]. Abstract: Basal transcription factors have never been considered as a priority target in the field of drug discovery. However, their unparalleled roles in decoding the genetic information in response to the appropriate signal and their association with the disease progression are very well-established phenomena. Instead of considering transcription factors as such a target, in this review, we discuss about the potential of the regulatory mechanisms that control their gene expression. Based on our recent understanding about the critical roles of c-MYB at the cellular and molecular level in several types of cancers, we discuss here how MLL-fusion protein centred SEC in leukaemia, ligand-estrogen receptor (ER) complex in breast cancer (BC) and NF-κB and associated factors in colorectal cancer regulate the transcription of this gene. We further discuss plausible strategies, specific to each cancer type, to target those bona fide activators/co-activators, which control the regulation of this gene and therefore to shed fresh light in targeting the transcriptional regulation as a novel approach to the future drug discovery in cancer. -
Supplementary Table 2
Supplementary Table 2. Differentially Expressed Genes following Sham treatment relative to Untreated Controls Fold Change Accession Name Symbol 3 h 12 h NM_013121 CD28 antigen Cd28 12.82 BG665360 FMS-like tyrosine kinase 1 Flt1 9.63 NM_012701 Adrenergic receptor, beta 1 Adrb1 8.24 0.46 U20796 Nuclear receptor subfamily 1, group D, member 2 Nr1d2 7.22 NM_017116 Calpain 2 Capn2 6.41 BE097282 Guanine nucleotide binding protein, alpha 12 Gna12 6.21 NM_053328 Basic helix-loop-helix domain containing, class B2 Bhlhb2 5.79 NM_053831 Guanylate cyclase 2f Gucy2f 5.71 AW251703 Tumor necrosis factor receptor superfamily, member 12a Tnfrsf12a 5.57 NM_021691 Twist homolog 2 (Drosophila) Twist2 5.42 NM_133550 Fc receptor, IgE, low affinity II, alpha polypeptide Fcer2a 4.93 NM_031120 Signal sequence receptor, gamma Ssr3 4.84 NM_053544 Secreted frizzled-related protein 4 Sfrp4 4.73 NM_053910 Pleckstrin homology, Sec7 and coiled/coil domains 1 Pscd1 4.69 BE113233 Suppressor of cytokine signaling 2 Socs2 4.68 NM_053949 Potassium voltage-gated channel, subfamily H (eag- Kcnh2 4.60 related), member 2 NM_017305 Glutamate cysteine ligase, modifier subunit Gclm 4.59 NM_017309 Protein phospatase 3, regulatory subunit B, alpha Ppp3r1 4.54 isoform,type 1 NM_012765 5-hydroxytryptamine (serotonin) receptor 2C Htr2c 4.46 NM_017218 V-erb-b2 erythroblastic leukemia viral oncogene homolog Erbb3 4.42 3 (avian) AW918369 Zinc finger protein 191 Zfp191 4.38 NM_031034 Guanine nucleotide binding protein, alpha 12 Gna12 4.38 NM_017020 Interleukin 6 receptor Il6r 4.37 AJ002942