analysis of signalling pathways by western blotting and immunoprecipitation
Tài liệu Báo cáo khoa học: Modulation of cyclin D1 and early growth response factor-1 gene expression in interleukin-1b-treated rat smooth muscle cells by n-6 and n-3 polyunsaturated fatty acids pdf
Ngày tải lên :
19/02/2014, 16:20
... synthesis and hyperphosphorylation of Rb by n-3 and n-6 PUFAs Induction of cyclin D1 is one of the earliest effects of mitogenic factors leading to cell cycle re-entry, G1-phase progression, and transition ... Action of EPA and DHA on cyclin D1 promoter activity induced by the Ras/Raf pathways Stimulation of Ras activity is known to promote cell cycle progression and a concomitant activation of cyclin ... the activity of the cyclin Ó FEBS 2004 Anti-mitogenic action of EPA and DHA (Eur J Biochem 271) 4469 D1 gene promoter is repressed by caveolin-1 [29] We decided to measure (by Western blotting) ...
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Báo cáo khoa học: Efficient inhibition of b-secretase gene expression in HEK293 cells by tRNAVal-driven and CTE-helicase associated hammerhead ribozymes doc
Ngày tải lên :
08/03/2014, 08:20
... level of extra- and intracellular release of b-amyloid peptide, which is a product of the catalytic activity of the target protein Our efforts to use Western blot analysis for detection of the ... b-amyloid peptide and dot blot analysis The level of b-amyloid peptide in conditioned cultured medium and in cells was determined by immunoprecipitation and dot blot analysis Conditioned medium ... preclearing process of the cell protein extract (100 lg of protein in 500 lL of extract) was carried out as described above Immunoprecipitation and determination of the intracellular level of Ab40 was...
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Báo cáo khoa học: Tumour necrosis factor-a attenuates insulin action on phosphoenolpyruvate carboxykinase gene expression and gluconeogenesis by altering the cellular localization of Foxa2 in HepG2 cells pptx
Ngày tải lên :
16/03/2014, 02:20
... was resolved by SDS-PAGE, transferred to poly(vinylidene difluoride) membranes and probed by western blotting with p-Akt and Akt (total) antibodies Each band is a representative of three independent ... presence of Foxa2 or Foxo1 by western blotting Each band is a representative of three separate blots from three independent experiments Blots were probed with TBP and b-actin antibodies and taken ... absence of any of these were taken as the control (Con) Two micrograms of total RNA were reverse transcribed with random primers and the levels of PEPCK and G6Pase mRNA were measured by RT-PCR...
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Báo cáo khoa học: Oxygen control of nif gene expression in Klebsiella pneumoniae depends on NifL reduction at the cytoplasmic membrane by electrons derived from the reduced quinone pool doc
Ngày tải lên :
17/03/2014, 10:20
... nitrogen-limited and anaerobic conditions and expression of NifL and NifA was induced from the tac promoter (Ptac) with 10 lM IPTG Expression of nifH¢-lacZ¢ was monitored by the determination of the b-galactosidase ... reduction of purified NifL using artificial electron donors or NADH and (b) analyzed the effect on nif-regulation of different membranebound oxidoreductases of the anaerobic respiratory chain and of terminal ... degradation of the FAD moiety The purified protein was dialyzed overnight into B buffer containing 25 mM potassium glutamate and used subsequently for biochemical analysis The amount of FAD cofactor of...
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Báo cáo y học: "Differential gene expression of bone anabolic factors and trabecular bone architectural changes in the proximal femoral shaft of primary hip osteoarthritis patients" pptx
Ngày tải lên :
09/08/2014, 08:23
... donors and donors' families for their kind donation of bone tissue used for this study and offer an extended appreciation to the orthopaedic surgeons and nursing staff of the Department of Orthopaedics ... of Orthopaedics and Trauma in the Royal Adelaide Hospital for support and cooperation in the collection of femoral specimens and to the mortuary staff of the Institute of Medical and Veterinary ... differential gene expression of the α1 and α2 chain of collagen type I (p < 0.0001 and p < 0.002, respectively) between OA and controls, are shown in Table Interestingly, the ratio of COL1A1/COL1A2 was...
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Báo cáo y học: " Profiles of cytokine and chemokine gene expression in human pulmonary epithelial cells induced by human and avian influenza viruses" doc
Ngày tải lên :
12/08/2014, 02:20
... of A/Thailand/1(KAN-1)/2004(H5N1) isolate; and Prof Malik Peiris for provision of A/Duck/Hong Kong/Y280/1997 (H9N2) isolate Author details Department of Microbiology, The Chinese University of ... There are qualitative and quantitative differences in the profiles of cytokines/chemokines induced by influenza viruses of different pathogenicity H5N1 was a more potent inducer of inflammatory cytokines/chemokines; ... tissue culture flasks for the inoculation of virus isolate at a multiplicity of infection (m.o.i.) of one After hour of adsorption, the virus was removed and ml of fresh RPMI-1640 media with 2% FBS,...
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Báo cáo y học: "Common gene expression strategies revealed by genome-wide analysis in yeast" ppt
Ngày tải lên :
14/08/2014, 08:20
... also found Because TLR is the product of TLRi and RA, this can be the result of the negative correlation of RA and RS and the lack of correlation between TLRi and RS However, no correlation Genome ... yeast, these different roles can be mirrored by the different behavior of protein and mRNA sub-profiles and, especially, by the different behaviors of RS and PS It seems that whereas PS works in ... blue) between RA and PA, PA and TLR or TLRi, RA and TR and between TR and TLR or TLRi Some of these correlations have been described previously [8,11,17,19] The correlation between TR and TLR was...
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Báo cáo sinh học: "Localization of leucocyte interferon gene in the q2.5 region of pig chromosomel by in situ hybridization" potx
Ngày tải lên :
14/08/2014, 19:23
... according to the G-band pattern, defined by the Committee for the standardized karyotype of the Domestic Pig (1988) 1986) The probe RESULTS AND DISCUSSION A total of 69 well-banded chromosome spreads ... 86% and the region is reduced from (q2.2 - q2.7) to one band q2.5 The number of grains pointed in this region is so high, that we could have reduced the number of spreads scored to one third and ... was labeled by the random primer method (Feinberg and Vogelmodified for tritium labeling The specific activity obtained equaled stein, 1983), H-labeled 10 dpm/pmg Appropriate amount of probe was...
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Báo cáo y học: "Differential gene expression in HIV/SIV-associated and spontaneous lymphomas"
Ngày tải lên :
02/11/2012, 11:08
... analysis of the transcription levels of the set, ND4, and SMG-1 genes in human spontaneous lymphomas and normal B- and Tlymphocytes (a) 32P-labeled PCR-fragments of the set and a-myb oncogenes, and ... the gene of protease calpain small subunit (capn4) and other (in case of lymphoma h1) and SMG-1, the gene of ribosomal protein S8 (in case of lymphoma h2), as well as several genes of unknown ... cDNA population of lymphoma h1 as tracer, and the cDNA population of lymphoma h2 as driver; and vice versa, 2) the cDNA population of lymphoma h2 as tracer, and the cDNA population of lymphoma h1...
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Báo cáo y học: "Transporter Molecules influence the Gene Expression in HeLa Cells"
Ngày tải lên :
03/11/2012, 11:44
... understanding of the influence of the molecular function is necessary, as well as of biochemical processes and of the biophysical properties provoked amongst others by a moderate repression of the ... RW1–buffer by centrifugation for 15 sec at 10000 rpm followed by addition of RPE-buffer × 500 µl and centrifugation at 10000 rpm for 15 sec Then the column was centrifuged to dryness and eluted by centrifugation ... block and the software version 1.2 The 10µl reaction consisted of 25ng of transcribed RNA (1µl), 1pmol Universal Probe (0.1µl), 50pmol each of forward + reverse primer (0.5µl), water (3.4µl) and...
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Ambient particulate air pollution induces oxidative stress and alterations of mitochondria and gene expression in brown and white adipose tissues ppt
Ngày tải lên :
06/03/2014, 19:20
... in the iBAT evaluated by Western blot A Representative bands of FA and PM2.5 on UCP1 protein level in iBAT by Western blotting B Quantitative results of Western blotting of UCP1 n = *P
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Báo cáo khoa học: Diverging regulation of pyruvate dehydrogenase kinase isoform gene expression in cultured human muscle cells pot
Ngày tải lên :
07/03/2014, 17:20
... dephosphorylation catalysed by pyruvate dehydrogenase phosphatase (PDP) To date, four isoforms of PDK (PDK1–4) and two isoforms of PDP (PDP1–2) have been identified in humans [3–5] These isoforms display unique ... human PDK1–4 and PDP1 were used in PCR and products were identified by gel electrophoresis (data not shown) Molecular cloning of each PDK or PDP isoform was confirmed by sequence comparison of each ... Regulation of hPDK2 and hPDK4 gene expression Table The effects of PPAR a, d and c agonists on PDK2 and PDK4 transcript abundance expressed as a percentage of basal levels Results are the means ± SEM of...
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Báo cáo khoa học: Gene expression in response to endoplasmic reticulum stress in Arabidopsis thaliana ppt
Ngày tải lên :
07/03/2014, 21:20
... two types of probes For analysis of differential gene expression, 1473 and 1703 beads were collected in fractions U1 and U2 of the up-regulated genes, and 3550 beads in fraction D of the down-regulated ... some of them might be missorted at the gates, which would expand the list of genes Of the beads collected at gates U1 and D on fluid microarray analysis, 89 and 87% were regarded as up-regulated and ... induced by ER stress by inhibiting BAX activation and translocation to mitochondria, by preserving the mitochondrial membrane potential, and by suppressing caspase activation [70] BAX and Bcl2, and...
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Báo cáo khoa học: Wild-type p53 enhances annexin IV gene expression in ovarian clear cell adenocarcinoma docx
Ngày tải lên :
22/03/2014, 16:20
... mRNA and protein levels of ANX4 and p21 were measured by western blotting and real-time RT-PCR analyses The p53 protein levels were also assessed by western blotting to verify that MMC and nutlin-3 ... Protein and RNA were extracted from two CCA (OVTOKO and OVISE) cells lines and one non-CCA (MCAS) cell line, and then ANX4 protein (A) and mRNA (B) levels were compared by western blotting and real-time ... Protein and total RNA were extracted from various EOC cell lines, HEK293 and LNCaP cell lines (A, B) Expression levels of protein and mRNA, and levels of p53, ANX4 and the known p53 targets, p21 and...
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Báo cáo khoa học: Differential gene expression in periportal and perivenous mouse hepatocytes potx
Ngày tải lên :
23/03/2014, 10:20
... expression in mouse liver separation of hepatocytes into periportal and perivenous subfractions was determined by real-time RT-PCR analysis of GS expression and western blotting for marker proteins as ... that several of the well-documented zonal differences in the levels and activities of key enzymes of various pathways of intermediary metabolism can be explained, at least in part, by corresponding ... nanograms of total RNA were reverse transcribed into cDNA by avian myeloblastosis virus-RT (Promega, Mannheim, Germany) using standard methods and oligo(dT)18 and random(dN)6 primers Expression analysis...
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Báo cáo khoa học: Effects of juvenile hormone on 20-hydroxyecdysone-inducible EcR, HR3, E75 gene expression in imaginal wing cells of Plodia interpunctella lepidoptera ppt
Ngày tải lên :
23/03/2014, 13:20
... B1 isoform) of other insect EcRs and a strong degree of identity was confined to the two ends of N-terminal region of the A/B domain Using a B1 isoform-specific probe from the A/B region of PIEcR, ... induction of mRNAs, the level of PIE75-B, PHR3 and PIEcR-B1 mRNAs was assessed after 1, and 18 h exposure, respectively Fig 8B and C show that after h and h exposure to 20E, the amount of accumulated ... effects of JH II on PHR3, PIEcR-B1, PIE75-B mRNAs induction by 20E were both dependent on the amount of JH II present and significant from 10)8 M JH II Fig Effect of 20E and JH II on induction of PIE75-B,...
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Báo cáo khoa học: Effect of ecdysone receptor gene switch ligands on endogenous gene expression in 293 cells docx
Ngày tải lên :
30/03/2014, 03:20
... days resulted in the death of 70% of the cells as indicated by observation of cell morphology To determine the possible genes and pathways that are affected by this ligand, we performed a microarray ... total of 20 lL reaction volume containing lL of cDNA, lL of each of forward and reverse sequence specific primers (from 10 lm primer stock), 10 lL of supermix (Bio-Rad Laboratories) and lL of nuclease ... affected by these ligands Only one or two genes have a cDNA sequence available for qRTPCR analysis and only half of the genes studied showed positive correlation between microarray and qRT-PCR analysis...
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Báo cáo hóa học: "Prognostic Impact of MiR-155 in Non-Small Cell Lung Cancer Evaluated by in Situ Hybridization" pot
Ngày tải lên :
18/06/2014, 16:20
... statistical analysis and drafted the manuscript KE, TB and KL carried out and supervised the ISH SWS and KE scored the cores KAS, SAS, SA and HS contributed in the clinical and demographic database and ... (SCC) and negative lymph node status (N0); (B) SCC and positive lymph node status (N+); (C) adenocarcinomas (AC) and N0; (D) AC and N+ Author details Department of Oncology, University Hospital of ... 2Institute of Clinical Medicine, University of Tromso, Tromso, Norway 3Department of Pathology, University Hospital of North Norway, Tromso, Norway 4Institute of Medical Biology, University of Tromso,...
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báo cáo hóa học:" Site-specific analysis of gene expression in early osteoarthritis using the Pond-Nuki model in dogs" pot
Ngày tải lên :
20/06/2014, 00:20
... μl of the 2X QuantiTect™ SYBR® green master mix, 0.1 μl of HK-UNG (Epicentre, Madison, WI), and μl of RNase-free water for each sample for a total volume of 20 μl The PCR profile consisted of ... observed in the HP content of the tissues between ACL-X and control joints for any of the regions tested Error bars indicate standard error of the mean Values are μg of HP/mg of tissue wet weight Differential ... observed in the GAG content of the tissues between ACL-X and control joints for any of the regions tested Error bars indicate standard error of the mean Values are μg of GAG/mg of tissue wet weight...
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