Detection of Viroid RNA and vd-siRNA in N. benthamiana Plants: Northern Blot Analyses for Viroid and vd-siRNAs

2021 ◽  
pp. 287-312
Author(s):  
Konstantina Katsarou ◽  
Nikoleta Kryovrysanaki ◽  
Kriton Kalantidis
Keyword(s):  
Northern Blot

RNA expression as a prognostic tool in idiopathic nephrotic syndrome

2007 ◽  
Vol 148 (23) ◽  
pp. 1067-1075
Author(s):  
Krisztina Fischer ◽  
Orsolya Galamb ◽  
Béla Molnár ◽  
Zsolt Tulassay ◽  
András Szabó
Keyword(s):  
Nephrotic Syndrome ◽  
Northern Blot ◽  
Idiopathic Nephrotic Syndrome ◽  
Minimal Change ◽  
Ribonuclease Protection Assay ◽  
Rna Expression ◽  
Rt Pcr ◽  
Protection Assay ◽  
Ribonuclease Protection

A gyermekkori nephrosis 90%-a idiopathiás nephrosis szindróma. Az idetartozó három kórkép, a minimal change betegség, a mesangialis proliferatio és a focalis sclerosis hasonló klinikai képpel jelentkező, eltérő prognózisú és terápiás válaszú betegség. Dolgozatunk célja az idiopathiás nephrosis szindrómába tartozó kórképek kialakulásával, progressziójával összefüggő genetikai ismeretek, génexpressziós változások áttekintése és funkcionális csoportosítása. A génexpressziós változások meghatározásának eszközeként, dolgozatunk röviden összefoglalja a northern blot, a ribonuclease protection assay, az in situ RNS-hibridizáció, a kvantitatív RT-PCR és a microarray módszerek lényegét. Az eddig elvégzett vizsgálatok a DNS-szintézis és repair gének, növekedési faktorok, extracelluláris mátrix, extracelluláris ligandreceptorok, extracelluláris jelátvitel zavarai mellett kiemelik a metabolikus és transzporter gének, illetve az immunszabályozó gének molekuláris eltéréseit, amelyek összefüggésben vannak az idiopathiás nephrosis szindróma eddig megismert molekuláris hátterével. A chiptechnológia fejlődésével és elterjedésével ezek a markerek és a hagyományos vizsgálati módszerek párhuzamos alkalmazása rutindiagnosztikai szempontból is fontossá válhat.

Northern Blot of tRNA in Yeast

BIO-PROTOCOL ◽  
2013 ◽  
Vol 3 (7) ◽  
Author(s):  
Yuehua Wei
Keyword(s):  
Northern Blot

scholarly journals Prediction, Microarray and Northern Blot Analyses Identify New Intergenic Small RNAs in Aliivibrio salmonicida

2012 ◽  
Vol 22 (6) ◽  
pp. 352-360 ◽  
Author(s):  
Rafi Ahmad ◽  
Geir Åsmund Hansen ◽  
Hilde Hansen ◽  
Erik Hjerde ◽  
Hege Lynum Pedersen ◽  
...  
Keyword(s):  
Northern Blot ◽  
Small Rnas ◽  
Aliivibrio Salmonicida

Expression and distribution of the Na+- HCO 3 − cotransporter in human pancreas

1999 ◽  
Vol 277 (2) ◽  
pp. G487-G494 ◽  
Author(s):  
Christopher R. Marino ◽  
Virginia Jeanes ◽  
Walter F. Boron ◽  
Bernhard M. Schmitt
Keyword(s):  
Northern Blot ◽  
Islet Cells ◽  
Rat Kidney ◽  
Physiological Role ◽  
Human Pancreas ◽  
Transmembrane Conductance Regulator ◽  
Transmembrane Conductance ◽  
Cellular Mechanisms ◽  
Double Labeling ◽  
Normal Human

The cellular mechanisms of [Formula: see text] secretion in the human pancreas are unclear. Expression of a Na+-[Formula: see text]cotransporter (NBC) mRNA has been observed recently, but the distribution and physiological role of the NBC protein are not known. Here we examined the expression and localization of NBC in human pancreas by Northern blot, immunoblot, and immunofluorescence microscopy. Rat kidney NBC probes detected a single 9.5-kb band by Northern blot. On immunoblots, two polyclonal antisera directed against different epitopes of rat kidney NBC identified a single ∼130-kDa protein. In cryosections of normal human pancreas, both antisera labeled basolateral membranes of large, morphologically identifiable ducts and produced a distinct labeling pattern in the remainder of the parenchyma. In double-labeling experiments, NBC immunoreactivity in the parenchyma colocalized with the Na+-K+pump, a basolateral marker. In contrast, NBC and cystic fibrosis transmembrane conductance regulator, an apical membrane marker, were detected within the same histological structures but at different subcellular localizations. The NBC antisera did not label acinar or islet cells. Our observations suggest that secretion of[Formula: see text] by human pancreatic duct cells involves the basolateral uptake of Na+and[Formula: see text] via NBC, an electrogenic Na+-[Formula: see text]cotransporter.

scholarly journals IL-8 and MCP Gene Expression and Production by LPS-Stimulated Human Corneal Stromal Cells

2012 ◽  
Vol 2012 ◽  
pp. 1-4 ◽  
Author(s):  
Roni M. Shtein ◽  
Susan G. Elner ◽  
Zong-Mei Bian ◽  
Victor M. Elner
Keyword(s):  
Gene Expression ◽  
Northern Blot ◽  
Stromal Cells ◽  
Blot Analysis ◽  
Time Course ◽  
Northern Blot Analysis ◽  
Statistical Significance ◽  
Dependent Manner ◽  
Corneal Inflammation ◽  
Chemotactic Protein

Purpose. To determine time course of effect of lipopolysaccharide (LPS) on production of interleukin-8 (IL-8) and monocyte chemotactic protein (MCP) by cultured human corneal stromal cells.Methods. Human corneal stromal cells were harvested from donor corneal specimens, and fourth to sixth passaged cells were used. Cell cultures were stimulated with LPS for 2, 4, 8, and 24 hours. Northern blot analysis of IL-8 and MCP gene expression and ELISA for IL-8 and MCP secretion were performed. ELISA results were analyzed for statistical significance using two-tailed Student'st-test.Results. Northern blot analysis demonstrated significantly increased IL-8 and MCP gene expression after 4 and 8 hours of exposure to LPS. ELISA for secreted IL-8 and MCP demonstrated statistically significant increases (P<0.05) after corneal stromal cell stimulation with LPS.Conclusions. This paper suggests that human corneal stromal cells may participate in corneal inflammation by secreting potent leukocyte chemotactic and activating proteins in a time-dependent manner when exposed to LPS.

Purification and characterization of the plasminogen activator inhibitors PAI-1, PAI-2, and PN-1 from the human glioblastoma U138

1993 ◽  
Vol 71 (5-6) ◽  
pp. 248-254 ◽  
Author(s):  
Patricia G. Murphy ◽  
Steven P. Lenz ◽  
Mark Dobson ◽  
Allan D. Arndt ◽  
David A. Hart
Keyword(s):  
Monoclonal Antibody ◽  
Plasminogen Activator ◽  
Concanavalin A ◽  
Northern Blot ◽  
Northern Blot Analysis ◽  
Plasminogen Activator Inhibitor ◽  
Immunoaffinity Column ◽  
Positive Hybridization ◽  
Activator Inhibitor ◽  
Pai 1

This investigation presents data which indicate that the plasminogen activator inhibitor (PAI) activity secreted from U138 cells is composed of three separate PAIs: PAI-1, PAI-2, and PN-1. It was demonstrated that the U138 PAI-1-like protein had an apparent molecular mass of 50 kilodaltons (kDa) and was purified to apparent homogeneity by elution from an anti-PAI-1 immunoaffinity column. These fractions were also reactive with a second anti-PAI-1 monoclonal antibody using immunoblotting techniques. Northern blot analysis of RNA isolated from unstimulated U138 cells demonstrated positive hybridization with the cDNA specific for human PAI-1. The U138 PAI-2-like protein was adherent to an anti-PAI-2 immunoaffinity column and was demonstrated to be nonadherent to concanavalin A – agarose, heparin–Sepharose, and the anti-PAI-1 immunoaffinity column. The eluted U138 PAI-2-like protein was demonstrated to have an apparent molecular mass of 60 kDa and was also reactive with a second anti-PAI-2 monoclonal antibody using immunoblotting techniques. Further, the cDNA specific for PAI-2 was demonstrated to hybridize to a 2.5-kilobase message from RNA isolated from U138 cells. A third PAI was detected that was nonadherent to concanavalin A – agarose and both of the anti-PAI columns. This 50-kDa PAI was adherent to heparin–Sepharose and thrombin–agarose columns, and was not reactive with any antibodies for either PAI-1 or PAI-2. Northern blot analysis of U138 RNA demonstrated positive hybridization with an oligodeoxynucleotide specific for PN-1. This investigation demonstrates with biochemical, immunological, and molecular data that the U138 glioblastoma constitutively produces three PAIs.Key words: plasminogen activator inhibitor, U138 glioblastoma, PAI purification, human tumor cell line, proteinase inhibitors.

scholarly journals Expression of tenascin in developing and adult mouse lymphoid organs.

1993 ◽  
Vol 41 (8) ◽  
pp. 1163-1169 ◽  
Author(s):  
G Ocklind ◽  
J Talts ◽  
R Fässler ◽  
A Mattsson ◽  
P Ekblom
Keyword(s):  
Lymph Nodes ◽  
Northern Blot ◽  
Adult Mouse ◽  
Histological Analysis ◽  
Lymphoid Cells ◽  
Differential Splicing ◽  
Cell Functions ◽  
Mouse Tissues ◽  
Adult Thymus ◽  
Short Splice Variant

The extracellular matrix (ECM) is essential in regulating many cell functions in non-lymphoid cells, and the ECM may also play a role in the function of the immune system. Tenascin is a hexameric glycoprotein of the ECM. In mouse, two major polypeptides of MW 210 KD and 260 KD are formed by differential splicing. Northern blot screening of various mouse tissues showed that the short 6 KB tenascin message was strongly expressed in the adult thymus, whereas very little or no tenascin mRNA could be detected in spleen. In addition, immunoblotting and histological analysis with monoclonal anti-tenascin antibodies revealed the presence of tenascin in lymph nodes and spleen. In thymus, only a short-splice variant of tenascin was detected by immunoblotting, which supported the Northern blot results. Immunohistology showed that the epithelial reticular stroma in both embryonic and adult mouse thymus expressed tenascin, as did the postnatal mesenchymal reticular stroma in lymph nodes and spleen. The distribution of tenascin in the thymus was more restricted than that of fibronectin and laminin.

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