dna damage response and mutagenesis in mouse embryonic stem cells pdf
Identification of oct4 and sox2 targets in mouse embryonic stem cells
... to the DNA via their respective DNA binding domains, and upon tethering of each factor to the enhancer region forming a ternary complex, new DNA- protein and protein-protein interactions induce ... Sox2 DNA interactions, characterized the joint DNA motif, expanded the Oct4 and Sox2 transcription binding network to the Smad1 and Stat3 signalling networks and utilized these binding data in ... Identification of Sox2 -DNA Interactions in Mouse Embryonic Stem Cells 5.1 Introduction 95 5.2 Results 96 5.2.1 Optimisation of ChIP and global mapping of Sox2 binding sites in mESCs 96 5.2.2 Sox2...
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Báo cáo y học: "Identification of a truncated form of methionine sulfoxide reductase a expressed in mouse embryonic stem cells" pps
... conditions in mouse embryonic stem cells Methods Mouse embryonic stem cell culture The mouse embryonic stem (MES) cells (CCE-24) were routinely grown on 0.1% gelatin-coated dishes in Dubecco’s ... AK was involved in manuscript writing and data analysis SL was involved in the conception and design, administrative support, data analysis and interpretation, manuscript writing and final approval ... hindering the access for the long form protein, thus having an advantage to be evolutionarily selected and maintained Since the truncated protein itself does not contain the c-terminal cysteines,...
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Báo cáo y học: "The impact of chromatin modifiers on the timing of locus replication in mouse embryonic stem cells" pot
... determined for retrotransposons (LINE and SINE B1) and rDNA repeats in wild-type OS25 ES cells and in mutant ES cells lacking Eed, Dnmt 1, Dnmt 3a/3b, G9a or Dicer The mean values and standard ... chromatin Other factors, including DNA methyltransferases, methyl -DNA binding proteins, polycomb repressor complexes (PRCs), nucleosome remodeling complexes and Dicer-dependent short interfering ... of DNA methylation and nuclear location (reviewed in [12]) Proteins capable of changing these parameters, chromatin modifiers, are important for establishing and maintaining particular chromatin...
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Dissecting transcriptional network in mouse embryonic stem cells
... medicine, disease modeling and drug development Notwithstanding these achievements, iPSCs technology remains in its infancy and a better understanding of the reprogramming process is required in ... genes in the list To check its expression pattern in both mouse and human ES cells and during differentiation, mouse E14 cells and human H1 cells were treated with retinoic acid (RA) to induce ... assays (EMSAs) – DNA binding domain of Zfp143 was cloned into pET42b (Novagen) The plasmid was transformed into BL21 E coli The DNA binding domain of Zfp143 protein was expressed and purified with...
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Báo cáo khoa học: The relative contribution of mannose salvage pathways to glycosylation in PMI-deficient mouse embryonic fibroblast cells pdf
... [18] and wild-type mouse embryonic fibroblast cells (WT cells) were grown in DMEM containing 20% fetal bovine serum CDG-Ib patient cells were grown in a-MEM containing 10% fetal bovine serum All media ... lower in CDG-Ib cells and decreased markedly in PMI-null cells However, CDG-Ib cells maintained sufficient glycosylation ability, and surprisingly, PMI-null cells showed only a slight reduction in ... A1-treated PMInull cells We thus performed mannose titration experiments in swainsonine-treated and bafilomycin A1-treated PMI-null cells Cells were treated with 10 lm swainsonine or 100 nm bafilomycin A1...
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Role of sonic hedgehog signalling in human embryonic stem cells and its neural derivatives
... culture feeding 3.2.3 Human embryonic stem cells and induced pluripotent stem cells Human embryonic stem cell line HES-3 (46, XX) was from ES Cell International (Singapore, http://www.escellinternational.com) ... forebrain, midbrain, hindbrain and the spinal cord This figure was reproduced from Epstein et al., 1999 2.4 SHH and neural development During the initial phase of neural induction, the ectoderm is induced ... effective in vitro system to study complex events underlying human nervous system development (Ben-Nun and Benvenisty, 2006; Dvash et al., 2006) 2.7 Embryonic stem cells and induced pluripotent stem cells...
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OCT 3 4 and SOX 2 are key factors for SDIA neurogenesis of mouse embryonic stem cells
... humor and inspiration, and providing invaluable thoughts on science and life, and all things in between Table of Contents Abstract Introduction Results Culturing of Mouse Embryonic Stem Cells (mESC) ... GFAP and Vimentin expressions in SDIA cultures List of Illustrations Illustration – Invitrogen BlockIT tetracycline inducible short, hairpin RNA system Introduction Mouse embryonic stem cells ... Sox-2 and Nanog have all been demonstrated to be involved in maintaining a pluripotent phenotype (Cavaleri and Scholer, 2003), the apparent inability of any single given factor to maintain mESC...
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Báo cáo khoa học: " Effect of dihydrotestosterone on mouse embryonic stem cells exposed to H2O2-induced oxidative stress" pdf
... nucleus and play a major role in triggering and coordinating gene responses [36] In addition, phosphorylation of JNK and p38 MAPK has been shown to play a role in cellular differentiation and inflammatory ... We determined the H2O2-induced ROS generation in mouse -5 embryonic stem cells using DCF-DA (10 M), a fluorescent probe of intracellular H2O2 Production of ROS was -3 detected in the cells exposed ... was loaded into a 96-well plate and examined using a luminometer (Victor3; PerkinElmer, Finland) and a fluorescent plate reader with excitation and emission wavelengths of 485 nm and 535 nm,...
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Báo cáo khoa học: " Radiosensitization by 2-benzoyl-3-phenyl-6,7-dichloroquinoxaline 1,4-dioxide under oxia and hypoxia in human colon cancer cells" pdf
... radio-sensitizer in hypoxic cells DCQ radiosensitization in the FHs74Int normal intestinal cell line After establishing effects of DCQ and IR in cancer cells, we compared DCQ efficacy in normal cells For ... fold in oxic cells and by 9.8 fold in hypoxic cells It is interesting to note that the significant inhibition of Cox-2 protein by DCQ in hypoxic and irradiated cells is associated with increased ... cancer cells confirms the greater radio-sensitizing effects of this drug in cancer cells DNA damage by DCQ in irradiated DLD-1 cells under oxia and hypoxia To determine if DCQ is a DNA- targeting...
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ROLE OF THE ERP29 IN MIGRATION, APOPTOSIS AND PROLIFERATION IN BONE MARROW STEM CELLS
... other cells by interacting with actin filaments in migrating cells Integrins can also act as signal transducer in the “inside-out signaling” model The activation of integrins is caused by talin binding ... profiling and ADF/cofilin, are also act as regulators in mediating actin polymerization by controling the availability of activated actin monomers, debranching and depolymerizing proteins and capping ... Protein kinase-like ER kinase PGE2 Prostaglandin E2 PI3K Phosphoinositide 3-kinase PTEN Phosphatase and tensin homolog PVDF Polyvinylidene difluoride Rb Retinoblastoma susceptibility protein RTN...
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Báo cáo khoa học: " Enhancement of radiosensitivity in human glioblastoma cells by the DNA N-mustard alkylating agent BO-1051 through augmented and sustained DNA damage response" potx
... design DNAdirected alkylating agents by linking the alkylating pharmacophore to the DNA- affinity molecules (e.g., DNA intercalating agents, DNA minor groove binder) [7,8] In most cases, the DNA- directed ... then washed in PBS twice, and the anti-gH2AX antibody was added at a dilution of 1:500 in 1% BSA and incubated overnight at 4℃ Again, the cells were washed twice in PBS before incubating in the dark ... produce DNA interstrand and/ or intrastrand cross-links [29,30] As has been known, bifunctional alkylating agents induce collapsed replication forks that can lead to either cell cycle arrest, DNA...
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Báo cáo y học: " Physical and in silico approaches identify DNA-PK in a Tax DNA-damage response interactome" pot
... certain types of damage induced in DNA, principally interstrand crosslinks [20] In contrast, Cluster includes key genes whose proteins mediate cell cycle arrest in response to genotoxic and other cellular ... arising from the original four Tax binding proteins known to be involved in the cellular DNA damage response There are five clusters with three bridge proteins; DNA- PK is one of the bridge proteins ... Tax-binding proteins generated from our physical mapping efforts, DNAPK was in the top five best represented binding proteins and occupied a top tier ranking via our functional clustering for DNA...
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Báo cáo khoa học: Dual role of Nbs1 in the ataxia telangiectasia mutateddependent DNA damage response ppt
... contains a forkhead-associated (FHA) domain and a Brca1 COOH-terminus (BRCT) domain in its NH2-terminal region, as well as an Mre11-binding domain and an ATM-binding domain in its COOHterminal ... checkpoint The Mre11-binding domain is responsible for binding to Mre11 during formation of the Mre11–Rad50–Nbs1 (MRN) complex The ATM-binding domain at the COOH-terminus binds to ATM and mediates ... 1) FHA and BRCT domains are often present in eukaryotic nuclear proteins involved in cell cycle checkpoint control or DNA repair The FHA domain appears to interact with target proteins in a phosphorylation-dependent...
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in situ detection of dna damage, methods and protocols
... Labeling DNA Damage with Terminal Transferase I LABELING DNA BREAKS USING TERMINAL TRANSFERASE (TUNEL ASSAY) Walker et al Labeling DNA Damage with Terminal Transferase Labeling DNA Damage ... DNA making it ideal for labeling fragments in apoptotic cells The enzyme will also label single stranded DNA molecules containing a 3'-OH and will attach nucleotides to a single-strand nick in ... methods for detecting DNA damage in situ is TdT-mediated dUTP-biotin nick end labeling (TUNEL) staining (1) TUNEL staining was initially described as a method for staining cells that have undergone...
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báo cáo hóa học:" Sodium arsenite and hyperthermia modulate cisplatin-DNA damage responses and enhance platinum accumulation in murine metastatic ovarian cancer xenograft after hyperthermic intraperitoneal chemotherapy (HIPEC)" pptx
... cancer cells were the kind gift of Dr Eddie Reed Cells were maintained in RPMI 1640 medium containing 10% fetal bovine serum, 100 μg/mL penicillin/streptomycin, mM L-glutamine and 0.2 units/mL insulin ... cisplatin -DNA damage involves the induction of DNA repair proteins to initiate DNA repair [10] We determined if NaAsO2 and hyperthermia modulated the expression of XPC, a platinum -DNA damage ... resulted in more Pt accumulating in the tumors when cisplatin is co-administered with sodium arsenite Cisplatin is a DNA damaging agent and p53 is implicated in platinum -DNA damage response [36]...
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Báo cáo khoa học: "Radiation induced apoptosis and initial DNA damage are inversely related in locally advanced breast cancer patients" ppt
... association between initial DNA damage, measured as DSB, and RIA Ionizing radiation (IR) kills cells by damaging the structure and function of genomic DNA The response of cells to this damage and their ... and has been involved in type of packaging likewise in the submission process PCL has been involved in conception and design of the study and in drafting the manuscript and has given final approval ... defined by two constants: the coefficient in origin a (as the origin of the curve in the Y axis determining the spontaneous apoptosis); and the coefficient b defining the slope of the curve a and...
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Tài liệu Báo cáo khoa học: Caveolin-1 influences P2X7 receptor expression and localization in mouse lung alveolar epithelial cells docx
... localization in subconfluent cells was determined Figure 5A shows a Caveolin-1 staining pattern at the cell surface, confirming the known caveolar location of Caveolin-1 In addition, intracellularly distinguishable ... localization and distribution of both proteins with a laser scanning confocal microscope, using two independent channels (Fig 5) Caveolin-1 and P2X7 receptor are endogenously expressed in E10 cells, and ... component In this study, the authors imply that P2X7 receptors in nonraft microdomains of submandibular gland cells are solely involved in ion channel activity, and that receptors present in lipid...
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báo cáo hóa học:" MicroRNA and gene expression patterns in the differentiation of human embryonic stem cells" doc
... reported in human [16,17] and mouse embryonic stem cells [18-20] The unique patterns of miRNA expression in embryonic stem cells suggest they are involved in maintaining "stemness" Identifying mRNAs ... groups: hES cells, EB, and adult cells Node A contained the genes that were upregulated in both hES cells and EB, node B contained the genes upregulated in hES cells only, node C contained the genes ... hES cells and adult cells was also significant (P < 0.05) In particular, the expression of miR-519b was 8-fold greater in hES cells than in EB cells and it was not even detected in adult cells...
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Báo cáo y học: " Developmental patterns and characteristics of epicardial cell markers Tbx18 and Wt1 in murine embryonic heart" ppsx
... published In this study, we employed immunofluorescence staining to investigate the expression of Tbx18 and Wt1 in heart using anti-Tbx18 and Wt1 antibodies and to identify whether Tbx18 and Wt1 ... expressed in the cardiomyocyte from E10.5 to at least E14.5 Interestingly, we found that, at E14.5, some Tbx18-expressing cells in the VS and LV did not colocalize with Nkx2.5, and Tbx18expressing cells, ... Tbx18 and Wt1 were expressed in the epicardial cells covering the heart after E10.5 (Figure 1, 2) Tbx18 began to be expressed in some cells within the VS and left ventricular wall at E10.5, and...
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Báo cáo y học: "Inflammatory responses in epithelia: endotoxininduced IL-6 secretion and iNOS/NO production are differentially regulated in mouse mammary epithelial cells" pot
... epithelial cells, likely to be the first cells in contact with infecting bacteria of the mammary gland, play an important role in innate immunity and in recruiting immune cells and orchestrating their ... forms, p65 and p50, increased their binding activity in response to ET compared to the basal binding activity in SCp2 cell nuclear protein extracts Figure 3A shows an abrupt increase in ET-induced ... tissues, and in the absence of immune cells The understanding of such innate immune responses of epithelia and the linkage thereof to chronic inflammation is crucial for understanding the link between...
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