Cell type specific applicability of 5-ethynyl-2'-deoxyuridine (EdU) for dynamic proliferation assessment in flow cytometry.
AuthorsDiermeier-Daucher S, Clarke ST, Hill D, Vollmann-Zwerenz A, Bradford JA, Brockhoff G,
JournalCytometry A
PubMed ID19235202
'Using the nucleoside analogue EdU (5-ethynyl-2''-deoxyuridine) for thymidine substitution instead of BrdU (5-bromo-2''-deoxyuridine) in cell proliferation assays has recently been proposed. However, the effect of EdU on cell viability, DNA synthesis, and cell cycle progression and consequently its usability for dynamic cell proliferation analysis in vitro has not been explored. ... More
CD133+ glioblastoma stem-like cells are radiosensitive with a defective DNA damage response compared with established cell lines.
AuthorsMcCord AM, Jamal M, Williams ES, Camphausen K, Tofilon PJ,
JournalClin Cancer Res
PubMed ID19671863
'PURPOSE: CD133+ glioblastoma tumor stem-like cells (TSC) have been defined as radioresistant. However, although previously classified relative to CD133- cells, the radiosensitivity of CD133+ TSCs with respect to the standard glioblastoma model, established glioma cell lines, has not been determined. Therefore, to better understand the radioresponse of this cancer stem ... More
TDAG51 is an ERK signaling target that opposes ERK-mediated HME16C mammary epithelial cell transformation.
AuthorsOberst MD, Beberman SJ, Zhao L, Yin JJ, Ward Y, Kelly K,
JournalBMC Cancer
PubMed ID18597688
'INTRODUCTION: Signaling downstream of Ras is mediated by three major pathways, Raf/ERK, phosphatidylinositol 3 kinase (PI3K), and Ral guanine nucleotide exchange factor (RalGEF). Ras signal transduction pathways play an important role in breast cancer progression, as evidenced by the frequent over-expression of the Ras-activating epidermal growth factor receptors EGFR and ... More
Click-iT assay with improved DNA distribution histograms.
AuthorsHamelik RM, Krishan A,
JournalCytometry A
PubMed ID19658154
'The Click-iT Assay developed and commercialized by Invitrogen is based on incorporation of a new 5-bromo-2''-deoxyuridine analog, 5-ethynyl-2''-deoxyuridine (EdU) into newly synthesized DNA and its recognition by azide dyes via a copper mediated ' ... More
Involvement of CD252 (CD134L) and IL-2 in the expression of cytotoxic proteins in bacterial- or viral-activated human T cells.
AuthorsWalch M, Rampini SK, Stoeckli I, Latinovic-Golic S, Dumrese C, Sundstrom H, Vogetseder A, Marino J, Glauser DL, van den Broek M, Sander P, Groscurth P, Ziegler U,
JournalJ Immunol
PubMed ID19494280
'Regulation of cytotoxic effector molecule expression in human CTLs after viral or bacterial activation is poorly understood. By using human autologous dendritic cells (DCs) to prime T lymphocytes, we found perforin only highly up-regulated in virus- (HSV-1, vaccinia virus) but not in intracellular bacteria- (Listeria innocua, Listeria monocytogenes, Mycobacterium tuberculosis, ... More
EdU, a new thymidine analogue for labelling proliferating cells in the nervous system.
AuthorsChehrehasa F, Meedeniya AC, Dwyer P, Abrahamsen G, Mackay-Sim A,
JournalJ Neurosci Methods
PubMed ID18996411
Labelling and identifying proliferating cells is central to understanding neurogenesis and neural lineages in vivo and in vitro. We present here a novel thymidine analogue, ethynyl deoxyuridine (EdU) for labelling dividing cells, detected with a fluorescent azide which forms a covalent bond via the ... More
EdU incorporation is an alternative non-radioactive assay to [(3)H]thymidine uptake for in vitro measurement of mice T-cell proliferations.
AuthorsYu Y, Arora A, Min W, Roifman CM, Grunebaum E,
JournalJ Immunol Methods
PubMed ID19647746
RATIONALE: T lymphocyte proliferations can be measured by [(3)H]thymidine incorporation. However, many labs avoid this technique because of the need to use radioactive substrates. In addition, [(3)H]thymidine incorporation method does not permit simultaneous characterization of the proliferating cells. We developed the 5-ethynyl-2'-deoxyuridine (EdU) and Cu(I)-catalyzed cycloaddition ... More
A chemical method for fast and sensitive detection of DNA synthesis in vivo.
AuthorsSalic A, Mitchison TJ,
JournalProc Natl Acad Sci U S A
PubMed ID18272492
We have developed a method to detect DNA synthesis in proliferating cells, based on the incorporation of 5-ethynyl-2'-deoxyuridine (EdU) and its subsequent detection by a fluorescent azide through a Cu(I)-catalyzed [3 + 2] cycloaddition reaction ("click" chemistry). Detection of the EdU label is highly sensitive and can be accomplished in ... More
p38 MAPK/MK2-mediated induction of miR-34c following DNA damage prevents Myc-dependent DNA replication.
AuthorsCannell IG, Kong YW, Johnston SJ, Chen ML, Collins HM, Dobbyn HC, Elia A, Kress TR, Dickens M, Clemens MJ, Heery DM, Gaestel M, Eilers M, Willis AE, Bushell M,
JournalProc Natl Acad Sci U S A
PubMed ID20212154
The DNA damage response activates several pathways that stall the cell cycle and allow DNA repair. These consist of the well-characterized ATR (Ataxia telangiectasia and Rad-3 related)/CHK1 and ATM (Ataxia telangiectasia mutated)/CHK2 pathways in addition to a newly identified ATM/ATR/p38MAPK/MK2 checkpoint. Crucial to maintaining the integrity of the genome is ... More
Detection of S-phase cell cycle progression using 5-ethynyl-2'-deoxyuridine incorporation with click chemistry, an alternative to using 5-bromo-2'-deoxyuridine antibodies.
The 5-bromo-2'-deoxyuridine (BrdU) labeling of cells followed by antibody staining has been the standard method for direct measurement of cells in the S-phase. Described is an improved method for the detection of S-phase cell cycle progression based upon the application of click chemistry, the copper(I)-catalyzed variant of the Huisgen [3+2] ... More
A rapid and robust assay for detection of S-phase cell cycle progression in plant cells and tissues by using ethynyl deoxyuridine.
AuthorsKotogány E, Dudits D, Horváth GV, Ayaydin F,
JournalPlant Methods
PubMed ID20181034
ABSTRACT: BACKGROUND: Progress in plant cell cycle research is highly dependent on reliable methods for detection of cells replicating DNA. Frequency of S-phase cells (cells in DNA synthesis phase) is a basic parameter in studies on the control of cell division cycle and the developmental events of plant cells. Here ... More
Induction of glutathione-dependent DNA double-strand breaks by the novel anticancer drug brostallicin.
AuthorsGuirouilh-Barbat J, Zhang YW, Pommier Y,
JournalMol Cancer Ther
PubMed ID19584235
Brostallicin is a DNA minor groove binder in phase II clinical trials. Here, we show that brostallicin induces gamma-H2AX nuclear foci that colocalize with 53BP1 and are dependent on glutathione, as shown by inhibition of those gamma-H2AX foci by l-buthionine sulfoximine. To differentiate brostallicin from the clinically approved minor groove ... More