linking rtks and anti bmp signals

... participated in study design and coordination and provided the expertise for RTPCR and withdrawal experiments HUS sub-cloned the PC12p75NTR(-) cells and participated in study design and result interpretation ... analysis (Top) and RT-PCR (Bottom) detecting iNOS protein and mRNA in total protein extracts and total RNA from PC12 cells simultaneously pre-treated for 24 hr with 10 ng/ml NGF and 10 ng/ml TNFα ... as mean +/standard error of the mean (S.E.M.), and analyzed by student unpaired two-tailed t test with significance set at p < 0.05 Results Combined NGF and TNFα induce iNOS message and protein...

Ngày tải lên: 19/06/2014, 22:20

... domains of Spir-2 and PTPN13 were generated by RT-PCR and inserted into pEGFP-C1 and pGEX-4T-2 The N- and C-terminal regions of the v-KIND KIND2 and the MAP2 CD were generated by PCR and cloned into ... immunostained with anti- Flag serum Flag immunoreactivity (red) and EGFP fluorescence (green) were observed by confocal microscopy Open and closed arrowheads indicate dendrites and axons, respectively ... immunoblotting (IB) with anti- v-KIND serum Immunoprecipitation assay (IP) of v-KIND from cerebellar lysates (input) by anti- MAP2 serum and normal mouse IgG as a positive and negative control, respectively...

Ngày tải lên: 14/02/2014, 19:20

... 5¢-GAGAATCCAGCTTCTTTCCC-3¢ and antisense, 5¢-GGCGACACTGTATGAATTGC-3¢); SENP2 (sense, 5¢-AACAGTCTCTACAATGCGGCCA-3¢ and antisense, 5¢-CCGTGTTCCATTACAAGCAGAA-3¢); SAE1 (sense, 5¢-GACCTGCTTCCCGATGACTTT-3¢ and antisense, ... 5¢-CAACAAAGAACCCTGATGGCACGA-3¢ and antisense, 5¢-GCATCCGTAGCTTGAACAAGCCTC-3¢); PIAS3 (sense, 5¢-ACTGCAGGGACCCTGCTACA-3¢ and antisense, 5¢-CTTGATCAGTGCTCGGGAATG-3¢); PIASxa (sense, 5¢-TGCACCTCATTCACCGTCAT-3¢ and antisense, ... adipose and may play a role in breast tumor development [13,14] Nuclear hormone receptors are typically ligand-activated transcription factors and their activities are modulated by coactivators and...

Ngày tải lên: 23/03/2014, 06:20

... collected in PBS and finally stored in cryoprotectant solution at -30°C Some of the brain sections were mounted on lysine-coated slides and stained with hematoxylin and eosin (H&E) Quantitative polymerase ... (H2O2) for 10 min, and incubated free-floating in antibodies (Abs) of polyclonal anti- mouse ionized calcium-binding adapter molecule (Iba-1, 1:1000; Wako, Osaka, Japan), monoclonal anti- mouse CD11b ... for NSE and S100 analysis in duplicate using available quantitative ‘sandwich’ enzyme-linked immunosorbent assay kits (Rapidbio, CA, USA) Sensitivity of the assays was 1.0 pg/ml for S100 and 0.1...

Ngày tải lên: 19/06/2014, 22:20

... (Figure 3A) and a reversible inhibition of both ERK (Figure 3B and C) and mTOR signaling (Figure 3D–H) Hence, resveratrol dose- and time-dependently activates AMPK and inhibits ERK and mTOR signaling ... contributions TJP, GD, DVT, OKM and MNA conceived of the study and designed experiments, DVT, OKM, MNA, NQ, MDF and TJP performed experiments, DVT, OKM and TJP analyzed data, DVT, OKM and TJP wrote the manuscript ... resveratrol at the time of incision and again day later showed blunted allodynia and this effect was dose-related (Figure 7A and B) Moreover, administering resveratrol and days following incision significantly...

Ngày tải lên: 21/06/2014, 19:20

... deparaffinized and hydrated in xylene and an ethanol series, post-fixed with 4% paraformaldehyde, and washed twice in phosphate-buffered saline For antigen retrieval in the detection of FRZB and β catenin, ... then washed twice in TBST, and incubated for hour with the secondary antibody For FRZB immunostaining, the secondary antibody was a biotin-conjugated rabbit anti- goat antibody (DAKO UK Ltd., Ely ... PS-1 antiserum [25] (a kind gift of P ten Dijke and C-H Heldin, Ludwig Institute for Cancer Research, Uppsala, Sweden) as primary antibody; as secondary antibody we used biotin-conjugated sheep anti- rabbit...

Ngày tải lên: 09/08/2014, 08:22

... nitrocellulose and probed with anti- phospho-ERK1/2 (1:2000, Sigma, M8159) or anti- phospho-Akt-Ser473 (1:1000, Cell Signaling, #9271) antibodies The blots were stripped and reprobed with antibodies ... incubated with anti- Flt-1 (sc-316), anti- Flk-1 (sc-504) or anti- Neuropilin (sc-5541) antibodies (2 µg/ml, Santa Cruz Biotechnology) in blocking buffer at 4°C o/n Unbound primary antibodies were ... serum, and [3H]thymidine incorporation was measured 72 hours after onset of stimulation The fluorometric CaspACE™ Assay System and western blot analyses with an anti- cleaved caspase-3 antibody...

Ngày tải lên: 11/08/2014, 08:20

... nitrocellulose membrane (Schleicher and Schuell, Dassel Germany) and probed with anti- phospho-Akt (Ser473) antibody (1:1000; Cell Signaling, Boston, MA, USA) or anti- armadillo antibody (1: 400; N2 7A1, ... categories and and the regulated Acknowledgements We thank Konrad Basler, George Hausmann and Thomas Radimerski for material and suggestion on signaling pathway activation, George Pyrowolakis and Roderic ... into cDNA and labeled with Cy5 or Cy3 fluorochromes; after hybridization of the cDNA to the microarray, the ratio of fluorescence between the Cy5 and Cy3 signals was measured, normalized and a Z-score...

Ngày tải lên: 14/08/2014, 21:20

... Biogenesis), CNTF (rabbit, Biogenesis) and GFAP (rabbit, DAKO) antiserum PB containing to 1% BSA and 0.3% Triton X-100 at 4℃ for a overnight As the kind of antiserum was not varied, double labeling ... immunoreactivity such as the substantia nigra and interpeduncular nucleus and to a lesser extent, the central gray matter (Fig 4E) The increase in the number and packing density of the GFAP-immunostained ... (C) In the pons, CNTF-IR neurons and glia were observed in the subtantia nigra and the facial nerves from PNW1 to PNW3 (D, E) Immunoreative developing neurons and astrocytes can be identified...

Ngày tải lên: 07/08/2014, 15:20

... the primers NGF-1F and NGF-1R and 400 bp with the primers NGF-2F and primer NGF-2R Overlapping PCR was performed to combine the PCR products of the partial dβ-NGF using NGF-1F and NGF-2R This overlap ... The phCMV vector and pcDNA2 were digested with Bgl II and EcoR I restriction enzymes, and purified following separation on a 1.5% agarose gel Ligation of the purified phCMV1 and pcDNA2 was performed ... (ELISA) development system (R&D Systems, USA) This system used a sandwich ELISA method with anti- human β-NGF as the detection antibody and was performed according to the manufacturer’s recommendations...

Ngày tải lên: 07/08/2014, 23:22

... insights into the cell types and cellular compartments, immunocytochemical methods were employed TGF-β isoforms 1, 2, and 3, TGF-β receptors I and II, Smads 2, 3, 4, 6, and 7, and phosphorylated Smad ... focus on members of the BMP family such as BMP- 2, BMP- 7, and growth differentiation factor (GDF-5) GDF-5 is also known as cartilage-derived morphogenetic protein (CDMP 1) and has been implicated ... Arthritis Research & Therapy Vol No Reddi R-Smads and the C-Smad have a highly conserved Mad homology domains (MH in the amino terminus and MH in the carboxy terminus) References Binding of...

Ngày tải lên: 09/08/2014, 07:20

... the SF cells are depicted in Table and outlined in detail by scatter plots in Figure (ligands) and Figure (receptors) As for the transcripts encoding the ligands, BDNF revealed high expression ... SD = standard deviation; SpA = spondyloarthritis In order to investigate the NGF/trkA-p75 axis as well as all other neurotrophic ligands and receptors at the transcript level, we used quantitative ... mainly by CD4+ T and B lymphocytes [1,21] Under inflammatory conditions such as allergy and arthritis, NGF can be produced, stored, and released by eosinophils, mast cells, lymphocytes, and synovial...

Ngày tải lên: 09/08/2014, 14:21

... degenerate and surgical degenerate NP samples Scatterplots were initially drawn to assess correlations between expression of MMP-10 and IL-1, TNF-α or NGF expression and between NGF and substance P, and ... paraformaldehyde/phosphate-buffered saline and processed into paraffin wax Five-micron sections from the tissue blocks were cut and stained with haematoxylin and eosin, and the degree of morphological degeneration ... PM degenerate tissues and -13 and ADAMTS-1, -4, -5, -9 and -15 [9,11,12,14,16,17] These studies have demonstrated significant increases in these enzymes during degeneration and have suggested vital...

Ngày tải lên: 09/08/2014, 14:22

... DMEM only, and stimulated cells were cultured in DMEM with cytokines tumor necrosis factor-alpha (TNF-α) (1 ng/mL) and interleukin-1-beta (IL-1β) (1 ng/mL) Data are expressed as mean ± standard deviation ... factor and receptor expression in rheumatoid arthritis and spondyloarthritis Arthritis Res Ther 2009, 11:R82 Raychaudhuri SP, Raychaudhuri SK: The regulatory role of nerve growth factor and its ... angiogenesis and cell trafficking [3] In patients with RA or PsA, pannus tissue adheres to the surface of articular cartilage; proliferating FLSs produce proteinases that degrade cartilage and underlying...

Ngày tải lên: 12/08/2014, 14:21

... tyrosine kinase signaling BMPs in lung development Several BMPs, including BMP3 , 4, and 7, are expressed during embryonic lung development [13,45,46] The expression of Bmp5 and Bmp7 has been detected ... and Smads 2, and are expressed in the adjacent epithelium The interactions of subsets of these ligand signals, particularly SHH, BMP4 and FGF10 have been extensively reviewed recently and several ... signaling pathway components, such as BMP receptors (type II and type I: ALK2, 3, and 6) and BMP specific receptor-regulated Smads (R-Smads), including Smad1, 5, and 8, are expressed in early mouse...

Ngày tải lên: 13/08/2014, 13:20

... sense and quantify changes in environmental salinity and to activate appropriate compensatory responses Thus, euryhaline fishes represent excellent models to identify and understand elements and ... molecular osmosensors that monitor and quantify environmental and extracellular osmolal- ity in fishes is minimal As is true for other cells, it is not clear how fish cells quantify osmolality to mount ... including shark rectal gland [12] and teleost gill and opercular membrane [13] Full-length transcripts of CaSR have been cloned from gilthead sea bream (Sparus aurata) [13] and spiny dogfish (Squalus...

Ngày tải lên: 18/02/2014, 16:20

... swelling stimulates anabolic metabolism and proliferation and provides cytoprotection, whereas cellular shrinkage leads to catabolism and insulin resistance and sensitizes cells to apoptotic stimuli ... [32], induction of the serum- and glucocorticoidinducible kinase Sgk [33], expression of the heat shock protein Hsp70 [34] and cyclooxygenase-2 [35], and a high antioxidant capacity [36] In hepatocytes, ... underlying the RVI, and thereby increase hepatocyte shrinkage by hyperosmolarity Mutual amplification of swelling and oxidative stress may be limited by the hepatocyte’s antioxidant and volume-regulatory...

Ngày tải lên: 18/02/2014, 16:20

... activation and platelet aggregation [18] We and others have shown that both the PI3-Kb and PI3-Kc isoforms contribute to the P2Y12-mediated stabilization of platelet aggregates under static and shear ... relies on both phospholipase C and PI3-K activity PI3-Kb and not PI3-Kc mediates the P2Y12 effect on thrombin-evoked Ca2+ responses In man and mouse, the PI3-Kb (p110b) and PI3-Kc (p110c) isoforms ... al tirofiban from Merck, Sharp and Dohme (Haarlem, the Netherlands); Akt inhibitor and LY294002 from Calbiochem (La Jolla, CA, USA) ET-18-OCH3, manoalide, U73122 and U73343 came from Biomol (Plymouth...

Ngày tải lên: 18/02/2014, 16:20

... (Welfide Corp., Osaka, Japan) Anti- CTGF/CCN2 sera have been described elsewhere [12,17] Anti- phospho-p38, anti- phospho-JNK, and anti- phospho-Akt/PKB were from New England Biolabs (Beverly, MA, USA) ... anisomycin and FBS on actin stress fibers in SMCs and their modulation by toxin B and latrunculin B Cells were first stimulated with either S1P, anisomycin or FBS for 30 and then fixed, permeabilized and ... characterized using muscle-specific antibodies against smooth muscle actin and myosin Cells were maintained in DMEM supplemented with 10% (v/v) fetal bovine serum (FBS) and antibiotics in a humidified...

Ngày tải lên: 19/02/2014, 16:20