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Blotting, Far-Western (0):
Recent article
Blotting, Far-Western
Biochemical Journal (427)3 2010
Abstract
We have sought to identify novel Golgi targets for mitotic kinases. We report in the present paper the identification of the ARF (ADP-ribosylation factor) exchange factor GBF1 (Golgi-specific brefeldin A-resistant guanine nucleotide-exchange factor 1) as a Golgi phosphoprotein. GBF1 is phosphorylate...
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PMID: 20175751
PDF is available here.
PDF is available here.
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Microbiology (155)Pt 7 2009
Abstract
We report that three (EF0089, EF2505 and EF1896, renamed here Fss1, Fss2 and Fss3, respectively, for Enterococcus faecalis surface protein) of the recently predicted MSCRAMMs (microbial surface components recognizing adhesive matrix molecules) in E. faecalis strain V583 bind fibrinogen (Fg). Despite...
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PMID: 19389755
PDF is available here.
PDF is available here.
Abstract
We have used affinity-purified GST-tagged mouse ERalpha-TAD fusion protein for interaction with nuclear proteins from the mouse brain. The pull-down assay and far-Western blotting detected four nuclear proteins of 100, 80, 68, and 50 kD. We have recently identified the 80-kD protein as MTA1 and demo...
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PMID: 19025768
PDF is available here.
PDF is available here.
Abstract
The three isoforms of the adaptor protein Shc play diverse roles in cell signalling. For example, the observation of p46 Shc in the nuclei of hepatocellular carcinoma cells suggests a function quite distinct from the better characterised cytoplasmic role. Ligands responsible for the transport of var...
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PMID: 19153664
PDF is available here.
PDF is available here.
Abstract
We here describe a rapid and simple protocol for far-western blotting, in which GST-tagged Src homology 2 (SH2) domains are used to probe cellular proteins in a phosphorylation-dependent manner....
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PMID: 19378071
PDF is available here.
PDF is available here.
Acta bio-medica de L'Ateneo parmense : organo della Societa di medicina e scienze naturali di Parma (80)3 2009
Abstract
HRPO can function as a partially-selective primary molecular probe when applied in either a conjugated or non-conjugated form. Some protein fractions can form complexes with HRPO through molecular mechanisms that involve physical interactions at the terminal alpha-mannose-rich regions of HRPO glycan...
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PMID: 20578419
PDF is available here.
PDF is available here.
Abstract
We propose two methods resolving this problem, either using new plasmids containing the Flag epitope, or using a trick to detect the T18 domain of adenylate cyclase. Furthermore, we describe a set of vectors for TAP, CBP or 6-histidine tagging that possess the same cloning site as our two-hybrid vec...
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PMID: 18924111
PDF is available here.
PDF is available here.
Abstract
We set out to identify novel Smad2 and Smad3 interacting proteins in epithelial cells. A non-transformed human cell line was transduced with Myc-His(6)-Smad2 or Myc-His(6)-Smad3-expressing retrovirus and was treated with TGF-beta1. Myc-His(6)-Smad2 or Myc-His(6)-Smad3 was purified by tandem affinity...
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PMID: 18729074
PDF is available here.
PDF is available here.
Infection and Immunity (76)10 2008
Abstract
We found that S. gordonii DL1 binds to HL-60 cells differentiated into monocytes, granulocytes, and macrophages. Using a glutathione S-transferase (GST) fusion to the NR2 domain, which is the sialic acid-binding region of Hsa, we confirmed that the Hsa NR2 domain also binds to differentiated HL-60 c...
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PMID: 18678668
PDF is available here.
PDF is available here.
Neuroreport (19)13 2008
Abstract
We investigated the expression of pannexins (Panxs). Western blot and reverse transcription-PCR detected the expression of Panx1 and Panx2. Immunolabeling localized Panx1 to the inner and outer sulcus, as well as the Claudius cells. Both Panx1 and Panx2 were expressed in the spiral and Scarpa's gang...
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PMID: 18695503
PDF is available here.
PDF is available here.
Journal of Bacteriology (19)13 2008
Abstract
We investigated the expression of pannexins (Panxs). Western blot and reverse transcription-PCR detected the expression of Panx1 and Panx2. Immunolabeling localized Panx1 to the inner and outer sulcus, as well as the Claudius cells. Both Panx1 and Panx2 were expressed in the spiral and Scarpa's gang...
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PMID: 18695503
PDF is available here.
PDF is available here.
Biochimica et Biophysica Acta (1783)8 2008
Abstract
We show that coexpression with H6PDH results in a dramatic shift from 11beta-HSD1 oxidase to reductase activity without affecting the activity of the endoplasmic reticular enzyme 17beta-HSD2. Immunoprecipitation experiments revealed coprecipitation of H6PDH with 11beta-HSD1 but not with the related...
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PMID: 18381077
PDF is available here.
PDF is available here.
Journal of Surgical Research (146)2 2008
Abstract
In this study, we showed for the first time the connection of ROR2 in Dupuytren's disease. ROR2 and myoglobin may play an important role in the pathophysiology of this disease....
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PMID: 17996904
PDF is available here.
PDF is available here.
Molecular Cell (28)6 2007
Abstract
We show that the DNA damage-inducible proteins UmuD(2) and RecA act in concert to modulate this mutagenic activity. Structural modeling suggests that the relatively open active site of DinB is enclosed by interaction with these proteins, thereby preventing the template bulging responsible for -1 fra...
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PMID: 18158902
PDF is available here.
PDF is available here.
FEBS Letters (581)23 2007
Abstract
p40(phox) activated phagocyte NADPH oxidase without p47(phox) in a cell-free system consisting of p67(phox), Rac and cytochrome b(558) relipidated with phosphatidylinositol 3-phosphate. The activation reached to 70% of that by p47(phox). Addition of p47(phox) slightly increased the activation, but n...
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PMID: 17803994
PDF is available here.
PDF is available here.
FEMS Microbiology Letters (274)1 2007
Abstract
Yeast two-hybrid and Far Western analyses were used to detect interactions between Bacillus subtilis spores' nutrient germinant receptor proteins and proteins encoded by the spoVA operon, all of which are involved in spore germination and located in the spores' inner membrane. These analyses indicat...
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PMID: 17573930
PDF is available here.
PDF is available here.
Liver International (27)7 2007
Abstract
These descriptive findings suggest an aberrant expression of Notch3 and Notch4 in HCC and allow the hypothesis of an activation of Notch signalling by Notch3....
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PMID: 17696940
PDF is available here.
PDF is available here.
Abstract
We propose that NbNACa1 is involved in BMV cell-to-cell movement through the regulation of BMV MP localization to the plasmodesmata....
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PMID: 17555275
PDF is available here.
PDF is available here.
Molecular Immunology (44)7 2007
Abstract
We investigated 10 male patients with XLA-compatible phenotype (agammaglobulinemia and undetectable B cells in peripheral blood) from 9 unrelated Central European families. We identified seven different mutations, six of which were novel. One previously described point mutation caused a premature st...
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PMID: 17045652
PDF is available here.
PDF is available here.
Genes & Development (21)2 2007
Abstract
We show here that mammalian Spt6 binds Ser2-phosphorylated (Ser2P) RNA polymerase II (RNAPII) through a primitive SH2 domain, which recognizes phosphoserine rather than phosphotyrosine residues. Surprisingly, a point mutation in the Spt6 SH2 domain (R1358K) blocked binding to RNAPIIo without affecti...
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PMID: 17234882
PDF is available here.
PDF is available here.
Abstract
Far western blotting (WB) was derived from the standard WB method to detect protein-protein interactions in vitro. In Far WB, proteins in a cell lysate containing prey proteins are firstly separated by SDS or native PAGE, and transferred to a membrane, as in a standard WB. The proteins in the membra...
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PMID: 18079728
PDF is available here.
PDF is available here.
Journal of Immunology (177)9 2006
Abstract
We previously showed that human NK cells used the NKp46 receptor to lyse Mycobacterium tuberculosis H37Ra-infected monocytes. To identify ligands on H37Ra-infected human mononuclear phagocytes, we used anti-NKp46 to immunoprecipitate NKp46 from NK cells bound to its ligand(s) on H37Ra-infected monoc...
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PMID: 17056548
PDF is available here.
PDF is available here.
Journal of Biological Chemistry (281)43 2006
Abstract
We have demonstrated that recombinant Ndk purified from wild-type E. coli contains significant UDG activity that is not intrinsic, but rather, is a consequence of a direct physical and functional interaction between Ung and Ndk, although a residual amount of intrinsic UDG activity exists as well. Co...
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PMID: 16895920
PDF is available here.
PDF is available here.
Molecular Microbiology (62)2 2006
Abstract
We show that RpfC, a protein consisting of N-terminal transmembrane, histidine kinase, response-regulator and C-terminal histidine phosphotransfer domains interacts with both RpfG, a protein consisting of an N-terminal response regulator domain and a C-terminal HD-GYP domain, and with RpfF. We also...
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PMID: 17020586
PDF is available here.
PDF is available here.
Molecular and Cellular Biochemistry (289)1-2 2006
Abstract
Our findings also suggested that the ankyrin D34 domain can interact directly with protein 4.2. The proper tertiary structures of these protein 4.2 fragments are essential for protein 4.2-ankyrin interaction. Meanwhile, ankyrin can enhance the interaction between protein 4.2 and CDB3....
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PMID: 16718373
PDF is available here.
PDF is available here.
Journal of Immunological Methods (313)1-2 2006
Abstract
We could trace the location of the epitopes on the HBsAg determinants. We also report the use of two specific methodology approaches based on molecular biology and biochemical techniques such as, respectively, cloning and expression of preS1 major neutralizing epitope of the HBsAg in Escherichia col...
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PMID: 16753174
PDF is available here.
PDF is available here.
Abstract
We show that overlay assay of bovine brain tubulin pool with glutathione-S-transferase (GST)-PLC-gamma1 PH domain fusion proteins, followed by matrix-assisted laser-desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), identified 68-kDa neurofilament light chain (NF-L) as a binding...
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PMID: 16819285
PDF is available here.
PDF is available here.
Journal of Immunology (176)12 2006
Abstract
We show that N. meningitidis binds fH to its surface. Binding to serogroups A, B, and C N. meningitidis strains was detected by FACS and Far Western blot analysis, and occurred in the absence of other serum factors such as C3b. Unlike Neisseria gonorrhoeae, binding of fH to N. meningitidis was indep...
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PMID: 16751403
PDF is available here.
PDF is available here.
Abstract
Myocilin forms high molecular weight complexes in vivo presumably due to interaction with itself and other myocilin binding proteins. To identify myocilin interacting proteins, yeast 2-hybrid analysis was performed on >1x10(6) human trabecular meshwork cDNA clones. Coimmunoprecipitation and Far West...
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PMID: 16289162
PDF is available here.
PDF is available here.
Abstract
The mammalian estrogen induced oviductal glycoprotein (OGP) has been known to associate with capacitated sperm, oocytes and developing embryos. This study aimed to identify the putative binding partner of OGP on gametes using N-terminal peptide of bonnet monkey (Macaca radiata) OGP, Nmon, as bait. A...
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PMID: 16567366
PDF is available here.
PDF is available here.
Zhonghua Yixue Zazhi (86)11 2006
Abstract
TGF-beta1 enhanced the FN mRNA expression in a dose- and time-dependent manner. The FN mRNA expression of the 5 ng/ml TGF-beta1 group was 3.7 times that of the control group (P < 0.05). The FN mRNA expression of the 15d-PGJ2, troglitazone-, and ciglitazone-pretreated groups were lower than that of t...
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PMID: 16681946
PDF is available here.
PDF is available here.
Abstract
We show here that 3H[A - 4] binds in a saturable fashion to the cell surface of C2C12 mouse fetal myoblasts at 4 degrees C, and not only binds at the surface but is internalized at 37 degrees C. "Far Western" immunohistochemistry performed on sections of E18 mouse incisors and molars with biotin-lab...
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PMID: 16214432
PDF is available here.
PDF is available here.
Journal of Molecular Biology (355)2 2006
Abstract
We utilized both genetic and surface plasmon resonance techniques to characterize the MLH1-PMS1-PCNA interaction. These analyses enabled us to determine the stability of the complex (K(D) = 300 nM) and to identify residues (572-579) in MLH1 and PCNA (126,128) that appear important to maintain this s...
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PMID: 16303135
PDF is available here.
PDF is available here.
Proteomics (6)1 2006
Abstract
We performed far-Western immunoblotting (protein overlay assay) to detect cellular PrP(C) interactors. Brain extracts of wild-type and PrP knockout mice were screened by far-Western immunoblotting for PrP-specific interactions. Subsequently, putative ligands were isolated by 2-DE and identified by M...
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PMID: 16294306
PDF is available here.
PDF is available here.
Journal of Infectious Diseases (192)12 2005
Abstract
Ligand-blotting and Biacore analyses revealed that the FbaA fragments harboring a proline-rich repeat domain (RD), but not the N- and C-terminal regions, possessed Fn-binding activity. Immunofluorescent microscopy findings showed that the N terminus and RD were exposed to external regions, which sug...
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PMID: 16288371
PDF is available here.
PDF is available here.
Biochimica et Biophysica Acta (1746)1 2005
Abstract
MUC7 is a low molecular weight monomeric mucin secreted by submandibular, sublingual and minor salivary glands. This mucin has been implicated in the non-immune host defense system in the oral cavity since it binds and agglutinates a variety of oral microbes. To investigate interactions between this...
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PMID: 16203048
PDF is available here.
PDF is available here.
Abstract
We examined were found to interact with their corresponding IcmQ proteins. Detailed bioinformatic, biochemical, and genetic analysis of the interaction between the variable FIR proteins and conserved IcmQ proteins revealed that their interaction depends on a variable region located between two conse...
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PMID: 16091472
PDF is available here.
PDF is available here.
Journal of Biological Chemistry (280)29 2005
Abstract
We found an interaction of TAK1 with all Smads tested, R-Smads Smads1-5, the co-Smad Smad4, and the inhibitory Smads (I-Smad6 and I-Smad7). Smad interaction with TAK1 takes place through their MH2 domain. This interaction is dependent on the presence of an active kinase domain in TAK1. TAK1 dramatic...
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PMID: 15911626
PDF is available here.
PDF is available here.
Nippon rinsho. Japanese journal of clinical medicine (63 Suppl 7) 2005
Abstract
Nippon rinsho. Japanese journal of clinical medicine (63 Suppl 7) 2005
Abstract
Abstract
We previously discovered a titin-like protein, smitin, associated with the contractile apparatus of smooth muscle cells. Purified native smooth muscle alpha-actinin binds with nanomolar affinity to smitin in smitin-myosin coassemblies in vitro. Smooth muscle alpha-actinin also interacts with striate...
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PMID: 15833278
PDF is available here.
PDF is available here.
Abstract
We used a 7-day-old rat model to elucidate the role of nuclear factor-kappaB (NF-kappaB) activation in HI stimulation of IL-1 signaling. HI was induced by permanent ligation of the left carotid artery followed by 90 min of hypoxia (7.8% O(2)). Using ELISA assays, we observed increased cell death and...
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PMID: 15773902
PDF is available here.
PDF is available here.
Cell Structure and Function (29)5-6 2005
Abstract
We searched for its binding proteins. Here we demonstrate the presence of two novel annexin-binding proteins. The homology search of partial amino acid sequences of these two proteins identified them as aminoacyl-tRNA synthetases (ARSs). Furthermore, antibody against aminoacyl-tRNA synthetases cross...
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PMID: 15840947
PDF is available here.
PDF is available here.
Abstract
Loss of buffalo Growth Hormone (buGH) in the various side fractions of standard buGH purification protocol has been determined quantitatively by direct binding ELISA and qualitatively by SDS-PAGE and Western blot analysis. Accounting result indicated that there was a considerable loss of buGH in the...
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PMID: 16239196
PDF is available here.
PDF is available here.
Abstract
We examined binding of Lf to Jurkat human lymphoblastic T cell line (Jurkat cells) by far Western blotting, and found that bovine Lf and human Lf bound to the same protein components of Jurkat cells, and that pepsin digestion of Lf disrupts the sites responsible for binding to cellular proteins. We...
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PMID: 15297761
PDF is available here.
PDF is available here.
Abstract
We examined the ability of T. brucei to bind bovine LF (bLF) by using a fluorescence test, streptavidin-biotin (SAB) microplate analysis, and far Western blotting using a biotin-streptavidin system. We found that bLF bound to components of T. brucei, and that bLF hydrolysate disrupted the sites resp...
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PMID: 15240935
PDF is available here.
PDF is available here.
Abstract
Methods in Enzymology (381) 2004
Abstract
Nucleic Acids Research (32)7 2004
Abstract
We have developed an efficient baculovirus expression system that allows us to obtain milligram quantities of recombinant RECQ1. Our gel filtration and dynamic light scattering experiments show that RECQ1 has an apparent molecular mass of 158 kDa and a hydrodynamic radius of 5.4 +/- 0.6 nm, suggesti...
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PMID: 15096578
PDF is available here.
PDF is available here.
Abstract
Abstract
Blot overlay is a useful method for studying protein-protein interactions. This technique involves fractionating proteins on SDS-PAGE, blotting to nitrocellulose or PVDF membrane, and then incubating with a probe of interest. The probe is typically a protein that is radiolabeled, biotinylated, or si...
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PMID: 15064457
PDF is available here.
PDF is available here.
Biochemical Journal (375)Pt 3 2003
Abstract
eIF2 (eukaryotic translation-initiation factor 2) is a substrate and an interacting partner for CK2 (protein kinase CK2). Co-immuno-precipitation of CK2 with eIF2beta has now been observed in HeLa cells, overexpressing haemagglutinin-tagged human recombinant eIF2beta. A direct association between Hi...
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PMID: 12901717
PDF is available here.
PDF is available here.
Abstract
We show that fusion complexes may exist as large soluble aggregates and thereby lead to false conclusions about the biological activity of RACK1. We developed a purification procedure that gave soluble monodisperse molecules of the protein. The RACK1 gene was cloned and expressed in a pMAL vector. A...
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PMID: 12963340
PDF is available here.
PDF is available here.
Abstract
We report here the analysis of a novel locus, blpGFEA, encoding four tandem paralogous proteins of approximately 30 kDa from B. hyodysenteriae. The four proteins share 31-39% sequence identity with lipoproteins from several species of bacterial pathogens, but the locus possesses a unique genetic org...
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PMID: 12706440
PDF is available here.
PDF is available here.
Abstract
We discuss alternative methods that exploit Src homology 2 (SH2) domains for profiling the tyrosine phosphoproteome. SH2 domains are small protein modules that bind specifically to tyrosine-phosphorylated peptides; there are more than 100 SH2 domains in the human genome, and different SH2 domains bi...
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PMID: 12754303
PDF is available here.
PDF is available here.
Journal of Biochemistry (133)2 2003
Abstract
We identified a calcineurin-binding protein in scallop testis by Far-Western blot analysis using purified calcineurin as a probe. The molecular mass of the binding protein estimated on the blot was 75 kDa. The isolated cDNA clone encoded a novel 474-residue protein, named CaNBP75. The region between...
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PMID: 12761177
PDF is available here.
PDF is available here.
Abstract
Abstract
Abstract
We demonstrated that PA has a unique chymotrypsin-like serine protease activity with Ser624 as an active site. To obtain further insight into the role of the protease activity of PA in viral proliferation, we examined the interaction between PA and matrix protein (M1). Both M1 purified from virion a...
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PMID: 12953845
PDF is available here.
PDF is available here.
European Journal of Biochemistry (269)18 2002
Abstract
We present data showing that p16 is a nuclear protein. First, after subcellular fractionation, p16 was clearly found in nuclei, although the protein is also present in other organelles. Immunocytochemical methods also confirm the above results. p16 seems to be firmly anchored to chromatin, as only e...
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PMID: 12230577
PDF is available here.
PDF is available here.