brdu staining and cell cycle analysis

... ELISA Sets and Kits for Quantitation of Analytes in Serum, Plasma, and Cell Culture Supernatants 143 Chapter 9: BrdU Staining and Multiparameter Flow Cytometric Analysis of the Cell Cycle ... For staining in tubes, wash the cells 1× with ml of staining buffer and pellet the cells by centrifugation (250 × g, min), and remove supernatant Resuspend and incubate cells in 100 µl of staining ... For staining in tubes, wash the cells 1× with ml of staining buffer and pellet the cells by centrifugation (250 × g for min), and remove supernatant Resuspend and incubate cells in 100 µl of staining...

Ngày tải lên: 11/04/2014, 10:34

... in cultured mouse cells p19ARF is predominantly upregulated during senescence by oncogene activation and/ or oxidative stress Senescence and Cell Cycle Control 265 In human cells undergoing replicative ... (1995) The role of Rb in cell cycle control Prog Cell Cycle Res 1:9–19 268 H Kiyokawa Hayflick L, Moorhead PS (1961) The serial cultivation of human diploid cell strains Exp Cell Res 25:585–621 Herbig ... suppression and cell mortality Cell 85:27–37 Serrano M, Lin AW, McCurrach ME, Beach D, Lowe SW (1997) Oncogenic ras provokes premature cell senescence associated with accumulation of p53 and p16INK4a Cell...

Ngày tải lên: 25/10/2013, 21:20

... not just in vitro and cell based but also simulated in silico Combining this method of analysis (in vitro, cell- based and in silico analysis) will facilitate systematic understanding of the underlying ... l-glutamine and 5% fetal bovine serum) and maintained at 37 C and 5% CO2 For translocation experiments, cells were removed with 0.05% trypsin EDTA, and plated with cell solution of ã 106 cellsặmL)1 ... prepared for cell assay and was added following the same procedure described for the time course cell assay The cells were washed with NaCl Pi and xed after 400 min, and the standard immunocytochemical...

Ngày tải lên: 16/03/2014, 11:20

... yeasts, cellsize checkpoints apparently operate to coordinate cell growth and cell- cycle progression For Schwann cells, by contrast, extracellular signals that stimulate cell growth, cellcycle ... absence of a cell- size checkpoint, because the larger cells not double their cell mass each cycle, and the smaller cells more than double their cell mass each cycle [10] Thus, proliferating cells can ... if large and small cells grow and progress through the cell cycle at the same rates (after Brooks [10]) The initial division is unequal and produces one cell of 10 mass units and one cell of mass...

Ngày tải lên: 06/08/2014, 18:20

... Cell cycle analysis of cells treated with or without drugs Cell cycle analysis of cells treated with or without drugs For fluorescence-activated cell sorting (FACS) analysis, both untreated and ... alternation in cell cycle progression to the proliferation of viruses HTLV-1 and/ or Tax-expressing cells have altered expression of some cell cycle- associated genes and accelerate cell cycle progression ... dysregulation of cell cycle regulatory proteins is considered to promote cell cycle progression and overcome cellular checkpoints Tax activates the expression of cyclin D2, cyclin E, CDK2, and CDK4 and the...

Ngày tải lên: 10/08/2014, 05:21

... rRNA Gluconeogenesis processing Stress TCA sub -cycle cell- cycle (G1) cell cycle G2/M cell cycle; M/G1 TCA Cell cycle Cell cycle Cell cycle sub -cycle (G1) (G2/M) (M/G1) Figure (see legend on previous ... of the basic cell cycle machinery in spore germination We used two cell cycle markers, the septin Cdc10 and the cyclin Clb2, to follow the timing and involvement of the mitotic cell cycle machinery ... are co-regulated during the mitotic cell cycle (Figure 6) Glucose induces events related to the cell cycle and advances the time of entering into the cell cycle upon subsequent transfer to rich...

Ngày tải lên: 14/08/2014, 08:20

... novel E2F-responsive and TRIP-Brregulated gene 88 FIGURE 19A: Cell cycle analysis of U2OS cells at 48 hours postdecoy peptide treatment 92 FIGURE 19B: Cell cycle analysis of U2OS cells at 72 hours ... U2OS cells 73 FIGURE 16C: FIGURE 16D: Inhibition of cell proliferation by decoy peptides 74 Cell cycle analysis of U2OS cells after decoy peptide 76 treatment xi FIGURE 17: Protein expression analysis ... (RT-PCR) 2.20 Cell Proliferation Assays 46 2.21 Determination of Cell Number 47 2.22 Colony Formation Assay 48 2.23 Cell Cycle Profile and TUNEL Staining Analyses by 48 Flow Cytometry 2.24 Cell Synchronization...

Ngày tải lên: 16/09/2015, 17:12

... Materials and Methods Chapter 2: Materials and methods 30 Chapter 2: Materials and Methods Chapter 2: Materials and methods 2.1 Routine cell culture of MN9D and SH-SY5Y cell lines The MN9D cell is ... gene and controls the cell cycle, S-phase, indirectly via a cellular cofactor in the murine cell line (Lee et al 1997) and nm23-M2 activates endogenous c-myc expression both at transcriptional and ... well as in many different cell lines cultured in vitro They have demonstrated that both Nurr1 and retinoids promotes cell cycle arrest at the G1 phase and induce a mature and highly differentiated...

Ngày tải lên: 08/11/2015, 17:09

... model and G1-control of the mammalian cell cycle Prog Cell Cycle Res 2000, 4:27-39 Cooper S, Shayman JA: Revisiting retinoblastoma protein phosphorylation during the mammalian cell cycle Cell ... 26:715-727 Cooper S: Cell cycle analysis and microarrays Trends Genet 2002, 18:289-290 Cooper S, Shedden K: Microarray analysis of gene expression during the cell cycle Cell Chromosome 2003, ... synchronized cells should be narrower than the size distribution of the original population, cell size should increase as the cells move through the cell cycle, and during the period of cell division...

Ngày tải lên: 13/08/2014, 23:20

... for controlling cell cycle progression (Morgan 1997; Zachariae and Nasmyth 1999) These studies and others demonstrated the universality of the cell Modeling Cell Cycle Control and Cancer with ... cell cycle machinery and the pRB and p53 tumor suppressor pathways The universality of the cell cycle in all eukaryotes strongly suggested that anticancer therapy aimed at regulators of the cell ... S-phase entry and cell cycle progression Overexpression of D- and E-type cyclins and Cdk4 is observed frequently in human tumors (see Sect 5), supporting the notion that these cell cycle regulators...

Ngày tải lên: 25/10/2013, 21:20

... most cell cycle responses While each single pocket protein can mediate cell cycle arrest upon cell- cell contact, pRb is the critical mediator of cell cycle arrest upon growth factor deprivation and ... in repression of E2F target genes and ultimately in a cell cycle arrest or exit from the cell cycle (see Fig 1) Thus, pRb can be viewed as a molecular cell cycle switch that is either turned ... delayed cell cycle exit of double knockout cells may indicate that oncogenic signals can induce a bypass of differentiation more easily in cells that lack both Rb and p107 or Rb and p130 In cells...

Ngày tải lên: 25/10/2013, 21:20

... mutant cells have been analysed phase of the cell- division cycle Therefore, we tested, using light microscopy and after DAPI staining, whether the slow growth yields a cell- cycle phenotype Cells ... the cell cycle and highly resistant to proteolytic degradation [40] Miscellaneous procedures Yeast were grown on standard media [41] and transformed as described by Gietz et al [42] Microscopy and ... in cell- cycle progression Yeast cells deprived of RIO1 lose minichromosomes at an increased rate relative to wild-type and arrest either as large G1 cells (i.e late in the G1 phase of the cell- division...

Ngày tải lên: 18/02/2014, 16:20

... emphasized by Adrian and Shin (2008), Brunnermeier and Pederson (2009), Kiyotaki and Moore (2009), Jermann and Quadrini (2009), Fostel and Geanakoplos (2009), Kurlat (2009) and others In the context ... Moral Hazard, Systematic Risk and Bailouts." mimeo, Harvard University and University of Toulouse [31] Fostel Ana and Geanakoplos, John and 2009, "Leverage Cycles and the Anxious Economy," American ... funds between a bank and non-financial firms in the same island In particular, we suppose that the bank is efficient at evaluating and monitoring non-financial firms of the same island, and also at enforcing...

Ngày tải lên: 06/03/2014, 08:20

... down-regulated upon cell cycle exit To this effect, HFF cells were deprived of serum for 48 h to induce cell cycle exit Serum was then re-added and samples taken as cells progressed into the cell cycle Total ... cell cycle progression, cells were incubated with BrdU 30 prior to fixation, and processed for BrdU staining as described in Materials and methods Percent of BrdU positive cells in each time point ... accumulation of BrdU positive cells Our data suggest that both Cdt1 and geminin are downregulated in quiescent HFF cells When cells re-enter the cell cycle, Cdt1 is expressed first, as cells prepare...

Ngày tải lên: 07/03/2014, 16:20

... PMLA induced a strain specific enhancement of cell growth and an equal (between strains) shortening of the cell cycle Materials and methods Strains and materials The following strains of P polycephalum ... 2A,C,E) and cell cycle duration between the third and fourth mitosis (Fig 2B,D,F) The ratio of cell sizes in the figures is expressed relative to control (mock ¼ water-injected) plasmodia and reflects ... which Ó FEBS 2004 Polymalate effects growth and cell cycle (Eur J Biochem 271) 3809 Fig Effects of polymalate injection on growth and duration of the cell cycle Variable amounts of PMLA (A–D) or poly(L-glutamate)...

Ngày tải lên: 16/03/2014, 18:20

... removed, and cells were washed several times with normal medium and then grown under standard conditions For proteasome inhibition, HeLa cells were synchronized by a double thymidine-block and released ... phases of the cell cycle, HeLa cells were arrested by a double-thymidine procedure at the G1 phase/S phase transition, and then released into the cycle after removing excess thymidine Cells were ... extracts were immunoprecipitated with specific antibodies and transferred to nitrocellulose membranes Immunostaining showed that cells in all cell cycle phases possess substantial amounts of chromatin-bound...

Ngày tải lên: 17/03/2014, 23:20

... during the cell cycle, we performed immunofluorescence staining of HeLa cells with anti-H3K4 trimethyl serum at different stages of the cell cycle The cell nucleus was counterstained and visualized ... a function of the cell cycle Western blot analysis of the whole -cell extract and histones from different stages of the cell cycle demonstrated that the overall levels of MLL1 and H3K4 trimethylation ... the cell cycle As MLLs and H3K4-specific methylations are well known to play critical roles in gene expression, we analyzed the dynamics and functions of MLLs and H3K4 methylation during cell cycle...

Ngày tải lên: 23/03/2014, 06:20

... lymphoma EL4 cells and 3T3 fibroblasts We analyzed the mRNA steadystate level and half-life of mouse cyclins and a selection of other cell cycle related genes for which important cell cycle- related ... elutriated cells The cell cycle distribution of the cell fractions is shown in Fig 5219 Cell cycle regulation of mouse cyclin mRNAs A Penelova et al Fig Cell cultures of mouse EL4 cell fractions ... vary during the cell cycle Cell Cycle 3, 1057–1061 Pines J & Hunter T (1989) Isolation of a human cyclin cDNA: evidence for cyclin mRNA and protein regulation in the cell cycle and for interaction...

Ngày tải lên: 30/03/2014, 20:20

... proliferation and motility and arrests Rat1/ras cell growth at the G1 phase of the cell cycle Experimental procedures Rat1/ras or Rat1/myr-ras cells was plated in agar at a density of 000 cells per ... and propidium iodide (PI, Sigma) as described before [38] A total of 10 000 stained cells were analysed in a fluorescence-activated cell sorter Materials and cell cultures Immunostaining All cell ... arrest of Rat1/ras cells at the G1 phase of the cell cycle We next examined the effect of HR12 on the distribution of Rat1/ras cells in the cell cycle To resolve the G1, S and G2/M phases, we...

Ngày tải lên: 31/03/2014, 01:20

... and Fission Yeasts Cell Cycle Molecules and Mechanisms of the Budding and Fission Yeasts Tim Humphrey and Amanda Pearce Summary The cell cycles of the budding yeast Saccharomyces cerevisiae and ... (MluI -cell cycle box) elements and is composed of Mbp1 and Swi6 MBF binding is cell cycle- regulated (reviewed in ref 12) Targets of MBF and SBF include cyclins, cell wall biosynthesis genes, and ... The Plant Cell Cycle 43 Modified Plant Cell Cycles Subheading described the typical cell cycle as observed in most meristematic and culture cells, but a number of variant types of cycle are found...

Ngày tải lên: 08/04/2014, 12:50