Abstract:
:A novel methodology for labeling, isolation, and detection of nucleic acids is described. Nucleic acid isolation is based on in vivo or in vitro incorporation of BrU or BrdU to either RNA or DNA, respectively, followed by immunoprecipitation of the labeled nucleic acid utilizing anti-BrdU MoAb, which crossreacts with BrU, attached to solid particles. Filter-bound bromine-labeled DNA or RNA was detected by immunoblotting with anti-BrdU MoAb, by a combined Southern/Western or Northern/Western approach, respectively. This method was applied to isolate and detect rRNA and mRNA from human cells, plasmid DNA from bacterial cells, and in vitro synthesized DNA. Newly transcribed BrU-labeled mRNA was recovered from the immunoprecipitates and analyzed by RT-PCR to study phorbol ester-mediated regulation of interleukin 1 gene transcription in human leukemic HL-60 or lymphoma U937 cells. The plasmid DNAs were isolated by immunoprecipitation from transformed bacterial cultures that were grown in the presence of BrdU and were detected immunochemically on filters. Likewise, the products of RT-PCR and Klenow polymerase-catalyzed DNA synthesis in which dTTP was replaced with BrdUTP were detected by immunoblotting. Since the method allows one to selectively separate or detect nucleic acids only synthesized during a pulse of the precursor, it can uniquely be used to identify nascent gene transcripts or the transcripts synthesized within specific time windows, e.g., after induction of differentiation, carcinogenesis, or drug treatment, and distinguish such transcripts from preexisting ones. In addition, this approach offers a simple and inexpensive alternative for preparing labeled DNA as well as RNA probes for use in a variety of hybridization protocols. Due to the low toxicity of BrU and BrdU, this approach can be used in analysis of gene transcription or DNA replication in vivo.
journal_name
Exp Cell Resjournal_title
Experimental cell researchauthors
Haider SR,Juan G,Traganos F,Darzynkiewicz Zdoi
10.1006/excr.1997.3644subject
Has Abstractpub_date
1997-08-01 00:00:00pages
498-506issue
2eissn
0014-4827issn
1090-2422pii
S0014-4827(97)93644-2journal_volume
234pub_type
杂志文章abstract::The centromere is a specialized region of the eukaryotic chromosome that is responsible for directing chromosome movements in mitosis and for coordinating the progression of mitotic events at the crucial transition between metaphase and anaphase. In this review, we will focus on recent advances in the understanding of...
journal_title:Experimental cell research
pub_type: 杂志文章,评审
doi:10.1006/excr.1998.4278
更新日期:1999-02-01 00:00:00
abstract::Hypoxia/reoxygenation (H/R) injury is characterized by microfilament reorganization in cardiomyocytes. Previous studies have shown that myofibrillogenesis regulator-1 (MR-1) is expressed in the myocardium and promotes actin organization in cardiomyocytes. The purpose of this study was to investigate the role of MR-1 i...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2015.05.026
更新日期:2015-10-01 00:00:00
abstract::Tubules and vesicles are membrane carriers involved in traffic along the endocytic and secretory routes. The small GTPase Arf6 regulates a recycling branch of short dynamic tubular intermediates used by major histocompatibility class I (MHC-I) molecules to traffic through vesicles between endosomes and the plasma memb...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2005.01.021
更新日期:2005-05-15 00:00:00
abstract::Using sequential extractions with buffers containing or not containing neutral detergent, two highly insoluble protein components were found in livers of 30-day-old rats. These compounds (molecular weights (MW) 52 900 and 33 200 respectively) were practically absent from livers of young rats (between 5 and 8 postnatal...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/0014-4827(84)90197-6
更新日期:1984-12-01 00:00:00
abstract::Plasmid 6.4 kbp DNA, 14 kbp DNA, lambda phage particles, all of which contained herpes simplex virus type 1 (HSV-1) thymidine kinase (TK) gene, or IgM molecules, were mixed with erythrocyte membranes and treated with neutral detergent. The transparent mixture was diluted with phosphate-buffered saline (PBS), followed ...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/s0014-4827(85)80014-8
更新日期:1985-08-01 00:00:00
abstract::Clear cell renal cell carcinoma (ccRCC) is one of the most common malignant tumors in the urinary system, and its incidence continues to increase. Regulator of calcineurin 1 (RCAN1), one of the genes on chromosome 21, is a crucial mediator of tumor inhibition. RCAN1.4 is best characterized as an endogenous inhibitor o...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2018.09.017
更新日期:2018-11-15 00:00:00
abstract::Synthetic tubugis are equally potent but more stable than their natural forms. Their anticancer potential was estimated on a solid melanoma in vitro and in vivo. Tubugi-1 induced the apoptosis in B16 cells accompanied with strong intracellular production of reactive species, subsequently imposing glutathione and thiol...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2019.04.028
更新日期:2019-07-15 00:00:00
abstract::Inhibition of cAMP-dependent protein kinase activity by microinjection of a specific physiologic protein inhibitor into sea urchin eggs inhibits the first cleavage after fertilization. Inhibition apparently occurs at some time prior to or during formation of the mitotic spindle. Measurement of the total protein kinase...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/0014-4827(90)90286-j
更新日期:1990-05-01 00:00:00
abstract::We had earlier demonstrated that a 46-kDa glycoprotein is involved in the differentiation of rat skeletal myoblasts. We now show that the binding of this glycoprotein to collagen and gelatin is disrupted by Arg-Gly-Asp (RGD) containing peptide but not by Arg-Gly-Glu (RGE). The former peptide also selectively elutes th...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/0014-4827(88)90368-0
更新日期:1988-11-01 00:00:00
abstract::Several cell division cycle (cdc) mutants of Saccharomyces cerevisiae (cdc28, cdc35, cdc19, cdc21, and cdc17) at the restrictive temperature (37 degrees C) in the presence of 1% glucose and defined medium divert most of the carbon (approximately 50%) to ethanol production with low biomass growth yields (Yglc) that cor...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1006/excr.1995.1062
更新日期:1995-03-01 00:00:00
abstract::Parietal yolk sac cells M1536 -B3 grown on cytodex 2 beads deposited an extracellular matrix on the surface of the beads. Cell-free matrix-coated beads were isolated by treatment of the cell monolayer with cytochalasin B (CB) at a concentration of 10 micrograms/ml of phosphate-buffered saline (PBS). The matrix when an...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/0014-4827(84)90652-9
更新日期:1984-06-01 00:00:00
abstract::After the 10th nuclear cycle the yolk centrosomes follow an irregular pathway. Unlike the somatic centrosomes, which move to the opposite poles of the nuclei to form the bipolar spindles, the yolk centrosomes remain as pairs at one pole of the yolk nuclei or shift feebly and nucleate irregular spindles, most of which ...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/0014-4827(91)90151-j
更新日期:1991-01-01 00:00:00
abstract::Mdg1/ERdj4, a mammalian chaperone that belongs to the HSP40 protein family, has been reported to be located in the endoplasmic reticulum (ER), is induced by ER stress, and protects ER stressed cells from apoptosis. Here we show that under normal physiological conditions, Mdg1/ERdj4 is expressed at various levels in th...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/s0014-4827(03)00316-1
更新日期:2003-10-15 00:00:00
abstract::Hypoxic blood-brain barrier (BBB) dysfunction is a common feature of CNS diseases however mechanisms underlying barrier disturbance are still largely unknown. This study investigated the role of transforming growth factor β (TGFβ), a cytokine known to induce expression of the proprotein convertase Furin, in hypoxia-me...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2019.111503
更新日期:2019-10-15 00:00:00
abstract::In studies of the ontogeny of fibroblast-epithelial interactions during late fetal lung rat lung development, we have identified two subpopulations of fibroblasts which differed in their ability to promote epithelial cell proliferation or differentiation. As glycosaminoglycans (GAGs) have been implicated in the regula...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/0014-4827(92)90072-g
更新日期:1992-10-01 00:00:00
abstract::p75NTR is a member of the tumor necrosis superfamily of proteins which is variably associated with induction of apoptosis and proliferation. Cyclin D2 is one of the mediators of cellular progression through G1 phase of the cell cycle. The present study demonstrates the inverse relationship between expression of cyclin...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2006.06.029
更新日期:2006-10-15 00:00:00
abstract::Human, Chinese hamster or Chinese hamster/human hybrid cytoplasts were fused with UV-irradiated xeroderma pigmentosum group A (XP-A) cells. Unscheduled DNA synthesis (UDS) of the XP-A nucleus was measured 0-2 and 2-4 h after seeding of the fused population. Human cytoplasts did correct the defect in the XP-A nucleus i...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/0014-4827(87)90209-6
更新日期:1987-04-01 00:00:00
abstract::Macrophage polarization plays a crucial role in regulating myocardial inflammation and injuries of coxsackievirus B3 (CVB3)-induced viral myocarditis (VM). It has been reported that miR-223 is a potent regulator of inflammatory responses that involved in macrophage polarization. However, the functional roles of miR-22...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2018.03.004
更新日期:2018-05-01 00:00:00
abstract::Statins are competitive inhibitors of hydroxy-methyl-glutaryl Coenzyme A (HMG-CoA) reductase, a key enzyme involved in the conversion of HMG-CoA to the cholesterol precursor mevalonate. Some statins, such as simvastatin (simvastatin), have been shown to have anti-cancer and anti-inflammatory effects, reducing cartilag...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2016.07.025
更新日期:2016-08-15 00:00:00
abstract::Skeletal muscles cope with a large range of activities, from being able to support the body weight during long periods of upright standing to perform explosive movements in response to an unexpected threat. This requires systems for energy metabolism that can provide energy during long periods of moderately increased ...
journal_title:Experimental cell research
pub_type: 杂志文章,评审
doi:10.1016/j.yexcr.2010.05.019
更新日期:2010-11-01 00:00:00
abstract::Polycystin-2 (PC2, TRPP2) is a nonselective cation channel whose dysfunction is associated with the onset of autosomal dominant polycystic kidney disease (ADPKD). PC2 contributes to Ca2+ transport and cell signaling in renal epithelia and other tissues. Little is known however, as to the external Ca2+ regulation of PC...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2016.11.004
更新日期:2017-01-01 00:00:00
abstract::Lithium has been found to be a novel inhibitor of the terminal differentiation of Friend murine erythroleukemia cells. A general method for the quantitative analysis of differentiation inhibitors has been developed and used to compare the site of inhibition by lithium with that by vanadate. Lithium inhibits the commit...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/0014-4827(87)90012-7
更新日期:1987-02-01 00:00:00
abstract::It has been generally accepted that alternatively activated macrophages (M2), which can be induced by type 2 cytokines such as IL-4, is responsible for tissue repair. However, the function of JNK in IL-4-induced M2 macrophage polarization remains unclear. Here, we demonstrated that M0 macrophages can be polarized into...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2017.05.010
更新日期:2017-08-15 00:00:00
abstract::Laminins are structurally and functionally major components of the extracellular matrix. Four isoforms of laminins (laminin-1, -2, -5 and -10) are expressed in a specific pattern along the crypt-villus axis of the intestine. Previous works indicated that expression of these isoforms is developmentally regulated and th...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2004.10.025
更新日期:2005-02-15 00:00:00
abstract::Endothelial cell-selective adhesion molecule (ESAM) is an immunoglobulin-like transmembrane protein associated with endothelial tight junctions (TJ). Based on a yeast two-hybrid screen, we have identified the membrane-associated guanylate kinase protein MAGI-1 as an intracellular binding partner of ESAM. MAGI-1 is a m...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2004.07.010
更新日期:2004-10-15 00:00:00
abstract::Skin extracellular matrix (ECM) molecules regulate a variety of cellular activities, including cell movement, which are central to wound healing and metastasis. Regulated cell movement is modulated by proteases and their associated molecules, including the serine proteases urinary-type plasminogen activator (uPA) and ...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1006/excr.2002.5644
更新日期:2002-11-01 00:00:00
abstract::Topoisomerase II is essential for cell proliferation and survival and has been a target of various anticancer drugs. ICRF-193 has long been used as a catalytic inhibitor to study the function of topoisomerase II. Here, we show that ICRF-193 treatment induces DNA damage signaling. Treatment with ICRF-193 induced G2 arr...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2006.02.029
更新日期:2006-07-01 00:00:00
abstract::Elongation factor-1 delta gene expression was analyzed during sea urchin development. EF-1 delta mRNA is present as a single 2.7-kb transcript in unfertilized eggs and in rapidly dividing cleavage stage embryos. It decreases rapidly 6 h after fertilization and then reappears at the gastrula stage as two transcripts of...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1006/excr.1998.4104
更新日期:1998-07-10 00:00:00
abstract:UNLABELLED:The success of hepatocyte transplantation has been limited by the low efficiency of transplanted cell integration into liver parenchyma. Human fetal hepatic progenitor cells (hepatoblasts) engraft more effectively than adult hepatocytes in mouse livers. However, the signals required for their integration are...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2009.07.007
更新日期:2009-11-15 00:00:00
abstract::Identification of transcription factors expressed by differentiated cells is informative not only of tissue-specific pathways, but to help identify master regulators for cellular reprogramming. If applied, such an approach could generate healthy autologous tissue-specific cells for clinical use where cells from the ho...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2018.05.031
更新日期:2018-08-15 00:00:00