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Blood, 1 December 2003, Vol. 102, No. 12, pp. 3938-3946. Prepublished online as a Blood First Edition Paper on August 7, 2003; DOI 10.1182/blood-2003-05-1479. This Article Full Text Full Text (PDF) All Versions of this Article: 2003-05-1479v1 102/12/3938    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Zhang, J. Articles by Lodish, H. F. Search for Related Content PubMed PubMed Citation Articles by Zhang, J. Articles by Lodish, H. F. Related Collections Hematopoiesis and Stem Cells Apoptosis Oncogenes and Tumor Suppressors Signal Transduction Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  HEMATOPOIESIS Role of Ras signaling in erythroid differentiation of mouse fetal liver cells: functional analysis by a flow cytometry–based novel culture system Jing Zhang, Merav Socolovsky, Alec W. Gross, and Harvey F. Lodish From the Whitehead Institute for Biomedical Research, Cambridge, MA; and the Department of Biology, Massachusetts Institute of Technology, Cambridge, MA. Ras signaling plays an important role in erythropoiesis. Its function has been extensively studied in erythroid and nonerythroid cell lines as well as in primary erythroblasts, but inconclusive results using conventional erythroid colony-forming unit (CFU-E) assays have been obtained concerning the role of Ras signaling in erythroid differentiation. Here we describe a novel culture system that supports terminal fetal liver erythroblast proliferation and differentiation and that closely recapitulates erythroid development in vivo. Erythroid differentiation is monitored step by step and quantitatively by a flow cytometry analysis; this analysis distinguishes CD71 and TER119 double-stained erythroblasts into different stages of differentiation. To study the role of Ras signaling in erythroid differentiation, different H-ras proteins were expressed in CFU-E progenitors and early erythroblasts with the use of a bicistronic retroviral system, and their effects on CFU-E colony formation and erythroid differentiation were analyzed. Only oncogenic H-ras, not dominant-negative H-ras, reduced CFU-E colony formation. Analysis of infected erythroblasts in our newly developed system showed that oncogenic H-ras blocks terminal erythroid differentiation, but not through promoting apoptosis of terminally differentiated erythroid cells. Rather, oncogenic H-ras promotes abnormal proliferation of CFU-E progenitors and early erythroblasts and supports their erythropoietin (Epo)–independent growth. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: T. Yamashita, O. Ohneda, A. Sakiyama, F. Iwata, K. Ohneda, and Y. Fujii-Kuriyama The microenvironment for erythropoiesis is regulated by HIF-2{alpha} through VCAM-1 in endothelial cells Blood, August 15, 2008; 112(4): 1482 - 1492. [Abstract] [Full Text] [PDF] T. Sawado, J. Halow, H. Im, T. Ragoczy, E. H. Bresnick, M. A. Bender, and M. Groudine H3 K79 dimethylation marks developmental activation of the {beta}-globin gene but is reduced upon LCR-mediated high-level transcription Blood, July 15, 2008; 112(2): 406 - 414. [Abstract] [Full Text] [PDF] F. Lohmann and J. J. 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