1β expression and action characteristics and pitfalls

... gene expression and gene expression of the senescence biomarker p16INK4a Seventeen NP samples were analysed for both caveolin-1 and p16INK4a gene expression using qRT-PCR Analysis of p16INK4a expression ... atherosclerotic patients and induced to senesce, caveolin-1 expression was correlated with senescence biomarkers and with expression of 4-hydroxynonenal expression (a marker of lipid peroxidation and thus oxidative ... examine caveolin-1 gene expression in human NP cells and to investigate any correlation between caveolin-1 and p16INK4a gene expression in isolated NP cells using the standard curve method of...

Ngày tải lên: 09/08/2014, 10:23

... protein and nucleic acids synthesis inhibition, depending on time and an infectious dose Despite the observed shutdown of host transcriptional and translational mechanisms and selective expression ... cDNA and 37 µl sterile water Reactions were carried out at the following conditions: 94°C for min, 60°C for min, and 72°C for 1.5 for 30 cycles (hsp70 and hsf1) or 95°C for min, 50°C for min, and ... the cells and mean values of at least 100 cells randomly selected on each sample were 24% and 46% for infected and control cells respectively HSP70 protein increases during infection and transiently...

Ngày tải lên: 11/08/2014, 08:21

... GPIIb-IIIa and CD107a expression (data not shown) Immunohistochemical analysis of P-selectin and PAI-1 expression In general, P-selectin and PAI-1 were expressed within the endothelium of arterioles and ... activation and temperature Statistics and graphics were performed using the software packages SigmaStat and SigmaPlot (Jandel Corporation, San Rafael, CA, USA) times of 255 ± 35 s and 238 ± 58 ... procoagulant state is not based on increased expression and release of P-selectin, Eselectin, ICAM-1, VCAM-1 and GPIIb-IIIa because tissue expression and plasma levels of these markers were not...

Ngày tải lên: 13/08/2014, 03:20

... Co-transfection of PKG1β and HIV-1 infectious molecular clone, pNL4-3, resulted in dose dependent increase in viral protein expression Expression of FLAG-tagged transfected PKG1β (top panel) and HIV-1 viral ... expression Figure Intact PKG1 β is required for activation of gene expression A) Schematic representations are shown of full length PKG1β (pCMV-PKG1β FLAG) and two deletion mutants, pCMV-PKG1β ... http://www.retrovirology.com/content/4/1/91 PKG1β activates expression from the HIV-1 LTR Figure PKG1β activates expression from the HIV-1 LTR A) Activation of HIV-1 LTR luciferase by PKG1β HeLa cells were cultured...

Ngày tải lên: 13/08/2014, 05:22

... my dad and mom; Winson and Cathryn, and sister; Gloria, thank you for always believing that I could achieve greater heights and to my beloved; Jonathan, for his endless support and understanding ... associated ROS and free radicals to the progression and development of neurodegenerative diseases such as schizophrenia and Alzheimer’s disease (AD) (Butterfield and Kanski, 2001; Rao and Balachandran, ... OH-, and OH) and RNS (i.e .NO and ONOO-) as it is metabolically active and contains a considerable amount of polyunsaturated fatty acids (PUFAs) that are highly vulnerable to peroxidation and...

Ngày tải lên: 01/10/2015, 17:27

... phosphorylated ERK, p38 and JNK (p-ERK, p-p38 and p-JNK) and total ERK, p38 and JNK (tERK, tp38 and tJNK) (B) The ratio of phosphorylated to total ERK (p44 ⁄ 42), p38 and JNK at each time point ... upregulates ICAM-1 expression in vivo and in vitro, and whether ERK, p38 or JNK, the MAPK family members, mediate LPS-induced ICAM1 expression in SCs We found that ICAM-1 expression in sciatic ... endothelial ICAM-1 expression and neutrophil adhesion J Immunol 162, 5466–5476 Smith CW, Marlin SD, Rothlein R, Toman C & Anderson DC (1989) Cooperative interactions of LFA-1 and Mac-1 with intercellular...

Ngày tải lên: 18/02/2014, 18:20

... cells and lymphatic endothelial cells in the lung [28,29]) As expected, Caveolin-1 and Flo-1 were predominantly found in the detergent-insoluble raft fraction (Fig 2A, second and third panels), and ... Flo-1 and P2X7 receptor in Triton X-100 E10 cells were lysed in a buffer containing 1% Triton X-100 to obtain soluble and insoluble fractions These fractions were adjusted to equal volumes, and ... fractions were collected (fraction 1, top of the gradient; fraction 13, bottom of the gradient), and an aliquot of each fraction (20 lL) was resolved by SDS ⁄ PAGE and subjected to western blot...

Ngày tải lên: 19/02/2014, 00:20

... Noxo1β Blot: anti-HA (+) Noxo1β (–) Noxo1γ PMA: Noxo1β C Noxo1γ B Noxo1β Fig Intracellular localization of Noxo1b and Noxo1c in CHO cells (A) Intracellular localization of Noxo1b (upper panels) and ... compilation ª 2006 FEBS 3673 Expression and function of Noxo1c R Takeya et al Expression of Noxo1b and Noxo1c in various human tissues The expression patterns of the Noxo1b and Noxo1c messengers were ... Noxo1b, Noxo1c, Nox1, and Noxa1 in various human cell lines: androgen-independent prostate cancer cells (LNCaP), androgen-independent prostate cancer cells (PC3 and DU145), and testicular germ...

Ngày tải lên: 19/02/2014, 06:20

... of the WISP1v proteins in HCS-2 ⁄ cells and their overexpression (A) Molecular constructs for the overexpression of WISP1, WISP1v and WISP1vx A mammalian expression vector, pFLAG-CMV was employed ... outcome of the constitutive expression of the full-length and the differentiation stage-dependent expression of WISP1v in vitro Effect of overexpression of CCN4/WISP1 and its variants on the phenotypes ... enhanced expression evaluated by real-time RT-PCR; ±, modest expression evaluated by real-time RT-PCR; –, expression undetectable ticular, expression of WISP1v among normal chondrocytes and osteoblastic...

Ngày tải lên: 07/03/2014, 10:20

... h for Western blot analysis of both ICAM-1 and GAPDH expression Representative blots are shown, and data (n ¼ 6) were normalized to GAPDH expression and are plotted as means ± SEM were stimulated ... only implicates the a and b isofroms of p38, as SB203580 has little or no effect on p38 c and d and it is possible that this could explain discrepancies between over -expression and inhibitor based ... inhibitors, PD098059 and U0126, were validated by analysis of phosphorylation of ERKs and 2, which lie downstream of MEK1 [24] Both TNFa and IL-1b upregulated phosphorylation of p42/44 ERKs and preincubation...

Ngày tải lên: 07/03/2014, 15:20

... later (a) HA-NBR1 (b) FEZ1-EGFP (c) CIB-myc protein expression, (d±f) coexpression of HA-NBR1 and FEZ1EGFP, and (g±i) coexpression of HA-NBR1 and CIB-myc The pattern of CIB staining observed in ... activation function, and thus could not be used to verify the NBR1/FEZ1 interaction, but the interaction between CIB and NBR1 was con®rmed Using the yeast two-hybrid assay, full length CIB and FEZ1 were ... To con®rm the interactions between NBR1, CIB and FEZ1, the full-length FEZ1 and CIB cDNAs were subcloned into the GAL4-BD vector pGBKT7 (Clontech) and tested in the yeast interaction assay with...

Ngày tải lên: 08/03/2014, 10:20

... with TBP and b-actin antibodies and taken as nuclear and cytosolic loading controls, respectively, and also used to ascertain the purity of the nuclear and cytosolic preparations (A) Bands were ... expression of rate-limiting enzymes of gluconeogenesis and glycogenolysis, including phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase), and by regulating glucagon and ... (ABI 7500, Applied Biosystems) Reactions were performed in triplicate and the expression of each transcript was quantified by the relative standard curve method and normalized to that of 18S rRNA...

Ngày tải lên: 16/03/2014, 02:20

... maturation, and the glycosylation was shown to affect integrin association and clustering, the capacity for fibronectin or lectin binding and the interaction with the regulatory gangliosides GT1b and ... read and translate such differences are the C-type lectins, siglecs and galectins [42] The ensuing microarray monitoring of expression of 42 C-type lectins and siglecs-2, -3, -5, -6, -7 and -9 ... Lensch M, Andre S & Gabius H-J (2006) Interaction profile of galectin-5 with free saccharides and mammalian glycoproteins: probing its fine specificity and the effect of naturally clustered ligand presentation...

Ngày tải lên: 16/03/2014, 10:20

... essential to elucidate their mode of action and their effects on the kinetic parameters of the 3Â-end processing and strand-transfer steps of the integration reaction Our results obtained in the ... the strand-transfer reaction When the effects of the various peptides on the strandtransfer step of the IN enzymatic reaction were studied, essentially the same pattern was observed (Fig and Table ... ) and no peptide (*)] IN strand-transfer activity was measured as described previously [58] and in Experimental procedures The Km and Vmax values for each IN peptide ratio were calculated and...

Ngày tải lên: 22/03/2014, 16:21

... p22phox and the Nox1 regulators NoxA1 and NoxO1 [5], but an increase in Nox1 gene expression is also essential Angiotensin II and platelet-derived growth factor lead to increased Nox1 mRNA levels and ... acquired and analyzed using an Olympus BX51 fluorescent microscope and DP Manager Software (Olympus, Japan) Cytosolic and nuclear fractionation Cells were plated in a 100 mm diameter dish and pretreated ... of macrophages and showed maximum effects at 45 with dose dependency (Fig 3A,B) To confirm the subcellular localization of b-catenin, we fractionated cells by cytosol and nuclear fraction after...

Ngày tải lên: 22/03/2014, 21:21

... basal promoter activity of KCTD10; and SP1 and AP-2a can regulate the promoter activity and endogenous KCTD10 expression oppositely through binding this region SP1 and AP-2a regulate KCTD10 Results ... [29]; (b) 1120 interaction model – the interaction between Sp1 and AP-2 affects formation of the Sp1–DNA complex [23]; (c) competing model – the AP-2 and Sp1 binding sites overlap, and the AP-2 binding ... Journal compilation ª 2009 FEBS R Liu et al SP1 and AP-2a regulate KCTD10 Roles of Sp1 and AP-2a in the regulation of the promoter activity and the expression of human KCTD10 Fig Transcription start...

Ngày tải lên: 30/03/2014, 02:20

... ischemia stimulates the expression of inflammatory cytokines (IL -1β, TNF-α), chemokines (IL-8, MCP1/CCL2) and adhesion molecules (ICAM-1) in the brain and in cultured astrocytes and brain endothelial ... transcriptional activation of MCP-1 and MCP-5 expression stimulated by hypoxia in human and mouse astrocytes, respectively Materials and methods Animal use and genotyping All procedures involving ... Media and cells were harvested for MCP-5 ELISA assay, RT-PCR detection of HIF-1α and MCP-5 mRNA expression, and western blot analysis of HIF-1α, respectively For FHAs, both hypoxic treatment and...

Ngày tải lên: 19/06/2014, 22:20

... cytokines and chemokines from primary brain pericytes constitutively and after LPS stimulation Brain pericytes were stimulated for 4, 8, and 24 h with LPS (0.1 and ug/ml) Media was collected and cytokine ... pericyte cultures with 0.1 and ug/ml LPS resulted in significant release of pro-inflammatory cytokines such as IL-1a, TNF-a, IL-3, IL-9 and IL-13 (4 h, h and 24 h) and anti-inflammatory cytokines ... nitric oxide and nitrosative stress in primary brain pericytes after LPS stimulation Brain pericytes were stimulated for 4, 8, and 24 h with LPS (0.1 and ug/ml), media collected, and analyzed...

Ngày tải lên: 19/06/2014, 22:20

... IL-8 expression in these patients in order to determine if IL-8 is upregulated in papillomas and whether defensin expression is associated with IL-8 expression We also attempted to relate IL-8 and ... hBD-3, HD5, HD6 and IL-8 mRNA expreshBD-1, hBD-2, hBD-3, HD5, HD6 and IL-8 mRNA expressions were analyzed by real-time RT-PCR Bars represent defensins expression normalized to β-actin and relative ... population, we did not identify a correlation between hBD expression and patient characteristics such as age, gender and racial background Expression of hBDs also appeared not to correlate with...

Ngày tải lên: 20/06/2014, 01:20

... affinity chromatography and characterization of the enzyme Material and methods Enzymes, chemicals and media Pfu+-polymerase was obtained from Roboclon (Berlin, Germany) and dNTPs from Roth (Karlsruhe, ... shifting the culture to 20°C and 200 rpm for another 16 h of cultivation Gene expression analysis Gene expression analysis was performed with crude cell extract Samples standardized to cell amount ... with 300 mM NaCl Fractions of washing and elution steps were collected to analyze purity by SDS-PAGE In order to remove imidazole and NaCl from the eluate, the pooled elution fractions were loaded...

Ngày tải lên: 21/06/2014, 02:20