flow cytometric analysis of kinase signaling cascades

... SLE Methodology of the analysis of MAPK and κ NF-κB activation by multiparameter intracellular flow cytometry Analysis of MAPK phosphorylation To validate flow cytometric analysis of MAPKs in human ... multiparameter flow cytometric analysis of signaling status of cells The flow cytometric technique allows simultaneous gating of multiple cellular subsets within a sample In addition, the percentage of cells ... protein kinases (MAPKs) Multiparameter intracellular flow cytometric analysis of STAT proteins and MAPKs Intracellular flow cytometric assays have been developed to assay general phosphorylation of...

Ngày tải lên: 09/08/2014, 01:23

... plants This type of data can be useful in planning molecular procedures for DNA polymorphism analysis or the study of genetic architecture of genomes by RFLP and PFGE (choice of rarely cleaving ... Galbraith DW, Harkins KR, Maddox JM, Ayres NM, Sharma DP, Firoozabady E (1983) Rapid flow cytometric analysis of the cell cycle in intact plant tissues Science 220, 1049-1051 Godelle B, Cartier ... (Provence) Three to four leaves of a single in vitro grown 2-month-old plantlet were chopped with a razor blade together with a leaf fragment of Petunia in mL of Marie’s nuclear isolation buffer...

Ngày tải lên: 08/08/2014, 18:21

... used for the analysis Results Evaluation of FCM analysis of GR In several tissues and cultured cells lines, administration of a GR agonist results in a significant down-regulation of GR This result ... monocytes (CD14+), Page of 11 (page number not for citation purposes) Arthritis Research & Therapy Vol 11 No Du et al Figure Evaluation of FCM analysis of GR binding by RLBA Analysis of GR binding in ... http://arthritis-research.com/content/11/4/R108 Figure Evaluation of FCM analysis of GR expression by western blot Analysis of GR repression in (left) K562 and (right) U937 with and without 10-6 M Dex of GR expression by western...

Ngày tải lên: 09/08/2014, 14:22

... range with a best straight line slope of 2.56, an intercept of 28.54 and a correlation coefficient of 0.996 The results of the analysis of some authentic samples of hydrogen peroxide are shown in ... evaluation gave a best straight line with a slope of 1.91, an intercept of 0.128 and a correlation coefficient of 0.998 The results of the analysis of some authentic samples (Table and Fig 6) show ... À 24.19), a slope of 4.91 and a correlation coefficient of 0.999 The analysis of some authentic samples of permanganate by this new method gave an average percentage recovery of 99.3% (Table and...

Ngày tải lên: 02/09/2015, 13:46

... the phosphorylation of the p46 isoform of JNK, whereas sorbitol treatment induced phosphorylation of both the p46 and p55 isoforms To confirm functional activation of the JNK signaling pathway, ... MAPK signaling, JSP1 was shown to be a positive regulator of the JNK signaling pathway [20,21] Since myristoylation determined the subcellular localization of JSP1, we asked whether abrogation of ... almost all of the existing cell lines we have examined not express detectable levels of JSP1 protein In fact, in order to generate sufficient quantities of wild-type protein for MS analysis of the...

Ngày tải lên: 16/02/2014, 14:20

... inhibits Matrigel invasion of these cells [33] TIMP-1 and TIMP-2 belong to a family of four different isoforms Of these, the prototype member of the TIMP family, TIMP-1, is of particular interest, ... Tiam1 ⁄ Rac-induced upregulation of tissue inhibitor of metalloproteinase-1 (TIMP-1) (A) Effects of catalase and specific inhibitors of mitogen-activated protein kinase kinase (MEK) 1,2 (PD098059, ... binding of nuclear extracts to the AP-1 site at )92 ⁄ )86 is reversed by catalase and by specific inhibition of ERK1,2 signaling To further substantiate the role of ROS and of ERK1,2 signaling...

Ngày tải lên: 19/02/2014, 05:20

... comparative analysis of signaling pathway(s) events in normal versus diseased tissue, (b) the comparative analysis of protein expression in various systems, (c) elucidation of the dynamic aspects of pathway ... modulation of regulatory networks [5,6] A direct benefit of such an approach would be the early recognition of ‘off target’ and side effects of drug candidates [7], as well as the identification of putative ... [11] Prominent players in these signaling cascades are protein kinases and phosphatases, which control the activation state of signaling pathways by the modulation of phosphorylation on Tyr, Ser...

Ngày tải lên: 16/03/2014, 00:20

... Intensity of cytoplasm Ratio of Translocation = Intensity of nucleus Intensity of cytoplasm Intensity of nucleus Scheme This is a simplication of a cell as seen by analysis software, where the software ... method of analysis (in vitro, cell-based and in silico analysis) will facilitate systematic understanding of the underlying properties of this signaling pathway To summarize Activation of cells ... analysis of NF-jB signaling pathway Tris HCl, pH 7.5, and 10 mm MgCl2 and typically included: 50 ng of rhIKK2; varying concentrations of SC-514 inhibitor [3000.1 lm; reconstituted 2.688 mg of...

Ngày tải lên: 16/03/2014, 11:20

... MKK kinase; MKK, MAPK kinase; MAPK, MAP kinase; TAK1, TGF-b-activated kinase 1; MEKK, MEK kinase; MLK, mixed lineage kinase; ASK1, apoptosis signal-regulating kinase 1; JNK, c-Jun N-terminal kinase ... Ó FEBS 2002 Regulation of SAPK signaling pathways (Eur J Biochem 269) 1061 Fig SAPK signaling modules The protein kinase cascades of SAPK signaling pathways and the points where ... the negative regulation of the MAP kinase signaling In the forthcoming paragraphs we present a detailed description of the mammalian DSPs involved in the regulation of SAPK signaling pathways MKP-1...

Ngày tải lên: 17/03/2014, 23:20

... expression of TH and DDC genes in MaBr4 cells (A) Western blots of ERK Phosphorylation of ERK in Mabr4 cells was observed after transfer from to 25 °C (B) RT-PCR analysis of expression of TH and ... after addition of GBP to the incubation medium of MaBr4 cells The involvement of a MAP kinase kinase (MEK) in the ERK activation pathway [19] was tested by evaluating the effect of U0126 U0126 ... in the presence of U0126 GBP clearly enhanced expression of both enzyme genes even in the presence of 10 lm U0126, indicating that the GBP signaling pathway for the stimulation of TH and DDC gene...

Ngày tải lên: 30/03/2014, 04:20

... Precision The precision of an analytical method describes the closeness of agreement (degree of scatter) between a series of measurements obtained from multiple sampling of the same homogenous ... number of IFNγ secreting cells (stimulated with gp100209, MART-127– 35, gp100pool, Flu, or PHA) – IFNγ secreting cells (stimulated with HIV as negative control) Statistical analysis Tetramer flow cytometric ... citation purposes) Journal of Translational Medicine 2008, 6:61 IFNγ Real Time PCR analysis was done using ABI Prism 7700 software for mRNA quantification Additional statistical analysis was performed...

Ngày tải lên: 18/06/2014, 15:20

... truncated forms of NS5A Expression and purification of N-terminally truncated forms of NS5A (a) Schematic of the structure of NS5A and the expressed truncated forms showing the locations of the N-terminal ... plasmon resonance analysis of the NS5A(Δ32):Fyn SH3 domain interaction SPR analysis was performed as described for NS5A(Δ32) and GST-FynSH3 The indicated concentrations of NS5A were flowed over a ... plasmon resonance analysis of the NS5A(Δ250):Fyn SH3 domain interaction SPR analysis was performed as described for NS5A(Δ250) and GST-FynSH3 The indicated concentrations of NS5A were flowed over a...

Ngày tải lên: 20/06/2014, 01:20

... and Paccierotti, F Flow cytometric assessment of trophosphamide toxicity on mouse spermatogenesis Cytometry 1996, 24, 174-180 10 Spano, M and Evenson, D P Flow cytometric analysis for reproductive ... for light protection of the propidium iodide After addition of solution C, the samples were kept in an ice bath for 30 to hours until analysis Flow cytometry The DNA contents of the dispersed testicular ... thereafter The proportion of diploid cells was 64% at the age of weeks, then decreased gradually through the age of 26 weeks The proportion of S-phase cells was also high at the age of weeks, then maintained...

Ngày tải lên: 07/08/2014, 14:23

... of cyclins versus total number of genes calculated with the number of proteins that match our cyclin HMMER profile bNumber of cyclins versus total number of genes calculated with the number of ... 4C) of the cells during the 24-h time series (b) Expression profiles of early cell cycle genes (c) Expression profiles of late cell cycle genes Error bars represent standard errors of the mean of ... the ability of the cells to recover from starvation (b) Expression profiles of early cell cycle genes at the time of sampling during the light experiment (c) Expression profiles of early cell...

Ngày tải lên: 09/08/2014, 20:21

... production of IFN-γ were rare Since the analysis of the combination of two functions decreases the background and the measurement of the IFN-γ+ MIP-1β+ T-cells was equivalent to the measurement of the ... is the method of choice for a detailed analysis of the immune response in a selected and limited number of samples In spite of an intense activity in the development and testing of new vaccines ... peptides Percentages of total IFN-γ+ and IFN-γ+ MIP-1β+ CD8 T-cells showed a goodness of fit of r2 = 0.9929 and a slope of 1.052 demonstrating an almost perfect linearity of the two measurements...

Ngày tải lên: 10/08/2014, 05:21

... pathways, such as mitogen-activated protein kinase growth signaling, causing inhibition of cell proliferation and induction of cell apoptosis [15,16] Analysis of the IκBα mRNA showed that bortezomib ... Pharmacological profile of SB 20 a selective inhibitor of cytokine suppressive binding protein/p38 kinase, in animal Page of (page number not for citation purposes) Journal of Inflammation 2005, ... expression and induction of IκBα in the IκBα-luc transgenic mice by LPS We demonstrated involvement of both the NF-κB and the p38 MAP kinase signaling pathways in the induction of IκBα expression by...

Ngày tải lên: 11/08/2014, 08:21

... fusion of tandem duplications of the gene or a mis-annotation of the genome or EST entry http://www.biomedcentral.com/1471-2229/8/1 Sequence analysis The majority of the overall protein sequence of ... biased removal of sequences and for a reproducible optimization of the sequence set, cluster analysis of sequences (CLANS) was used [52,53] The conserved region of the 811 hits of iteration was ... ancestral developmental tool kit of land plants International Journal of Plant Sciences 2007, 168(1):1-35 Arabidopsis Genome Initiative: Analysis of the genome sequence of the flowering plant Arabidopsis...

Ngày tải lên: 12/08/2014, 05:20

... change of the probe due to the binding of cyclic AMP was analysed in terms of (1) time lag between the direct addition of drug into the buffer surrounding the cell and the initiation of a measurable ... binding of cyclic AMP to Epac and a subsequent conformational change of Epac leading to an increased distance between the CFP and YFP termini of the probe and a subsequent loss of FRET The trace of ... differences in rate of onset of action of these drugs we explored the rate of cyclic Billington and Hall Respiratory Research 2011, 12:89 http://respiratory-research.com/content/12/1/89 Page of 10 Figure...

Ngày tải lên: 12/08/2014, 13:22

... The flow profiles and needle tip effects were examined using CFD for access flows of 400, 800, and 1200 mL/min with pump on and off at pump rates of 400 mL/min in the center of the lumen and off ... pump off and pump on, respectively Solving for the access flow QA gives the quadratic model: QA = QB/(1 - √(Pon/Poff)), (5) where QA depends on QB and the square root of the ratio of Pon and Poff ... steady flow models, which are first-order approximations to pulsatile flow One of the best described solutions for laminar flow through a straight circular tube of constant cross section is Page of...

Ngày tải lên: 13/08/2014, 16:21

... profile and the profiles of the CT-dependent EGF responsive gene group that was enriched for c-Ets1 related binding sites (p = × 10-4); between the C/EBPα expression profile and the profiles of ... established targets of EGFR signaling, two of them (Rorα and Rev-erb-alpha) are essential components of the circadian clock gene network in the SCN [22,38,39] Involvement of Rorα binding sites ... The signaling pathways downstream of EGFR are uncharacterized in the SCN, but by analogy to other wellcharacterized SCN inputs [5], and studies in other systems of context-dependent EGFR signaling...

Ngày tải lên: 14/08/2014, 16:21