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PMC:3317373 / 5456-7257 JSONTXT 9 Projects

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Id Subject Object Predicate Lexical cue
T2309 0-4 Sentence denotes 2.4.
T59 0-4 Sentence denotes 2.4.
T2310 5-48 Sentence denotes Electrophoretic Mobility Shift Assay (EMSA)
T60 5-48 Sentence denotes Electrophoretic Mobility Shift Assay (EMSA)
T2311 49-135 Sentence denotes Monolayers of astrocytes were washed with PBS and harvested by scraping into cold PBS.
T61 49-135 Sentence denotes Monolayers of astrocytes were washed with PBS and harvested by scraping into cold PBS.
T2312 136-328 Sentence denotes The cell pellet obtained by centrifugation was resuspended in buffer containing 10 mM HEPES (pH 7.9), 10 mM KCl, 0.1 mM EDTA, 0.1 mM EGTA, 1.0 mM DTT, and 0.5 mM phenylmethylsulfonyl fluoride.
T62 136-328 Sentence denotes The cell pellet obtained by centrifugation was resuspended in buffer containing 10 mM HEPES (pH 7.9), 10 mM KCl, 0.1 mM EDTA, 0.1 mM EGTA, 1.0 mM DTT, and 0.5 mM phenylmethylsulfonyl fluoride.
T2313 329-430 Sentence denotes Then 10% Nonidet P-40 was added and vortexed briefly, and the nuclei were pelleted by centrifugation.
T63 329-430 Sentence denotes Then 10% Nonidet P-40 was added and vortexed briefly, and the nuclei were pelleted by centrifugation.
T2314 431-604 Sentence denotes The nuclear proteins were extracted with buffer containing 20 mM HEPES (pH 7.9), 0.4 mM NaCl, 1.0 mM EDTA, 1.0 mM EGTA, 1.0 mM DTT, and 1.0 mM phenylmethylsulfonyl fluoride.
T64 431-604 Sentence denotes The nuclear proteins were extracted with buffer containing 20 mM HEPES (pH 7.9), 0.4 mM NaCl, 1.0 mM EDTA, 1.0 mM EGTA, 1.0 mM DTT, and 1.0 mM phenylmethylsulfonyl fluoride.
T2315 605-750 Sentence denotes Insoluble material was removed by centrifugation at 14000 rpm, and the supernatant containing the nuclear proteins was stored at –80°C until use.
T65 605-750 Sentence denotes Insoluble material was removed by centrifugation at 14000 rpm, and the supernatant containing the nuclear proteins was stored at –80°C until use.
T2316 751-908 Sentence denotes Protein concentration was determined using a bicinchoninic acid assay kit with bovine serum albumin as the standard (Pierce Biochemicals, Rockford, IL, USA).
T66 751-908 Sentence denotes Protein concentration was determined using a bicinchoninic acid assay kit with bovine serum albumin as the standard (Pierce Biochemicals, Rockford, IL, USA).
T2317 909-985 Sentence denotes EMSA was performed using gel shift assay system (Promega, Madison, WI, USA).
T67 909-985 Sentence denotes EMSA was performed using gel shift assay system (Promega, Madison, WI, USA).
T2318 986-1143 Sentence denotes Consensus oligonucleotide probe (5′-AGTTGAGGGGACTTTCCCAGGG-3′) was end-labeled by T4-polynucleotide kinase using [γ-32P]-ATP (Free Biotech., Beijing, China).
T68 986-1143 Sentence denotes Consensus oligonucleotide probe (5′-AGTTGAGGGGACTTTCCCAGGG-3′) was end-labeled by T4-polynucleotide kinase using [γ-32P]-ATP (Free Biotech., Beijing, China).
T2319 1144-1350 Sentence denotes Nuclear protein (20 μg) was preincubated in 20 μL binding buffer containing 10 mM Tris-HCl (PH 7.5), 1 mM MgCl2, 0.5 mM NaCl, 4% glycerol, 0.5 mM EDTA, 0.5 mM DTT, and 2 μg poly dI-dC for 20 minutes on ice.
T69 1144-1350 Sentence denotes Nuclear protein (20 μg) was preincubated in 20 μL binding buffer containing 10 mM Tris-HCl (PH 7.5), 1 mM MgCl2, 0.5 mM NaCl, 4% glycerol, 0.5 mM EDTA, 0.5 mM DTT, and 2 μg poly dI-dC for 20 minutes on ice.
T2320 1351-1463 Sentence denotes After addition of the 1 μL 32P-labled oligonucleotide probe, the incubation was continued for 20 minutes on ice.
T70 1351-1463 Sentence denotes After addition of the 1 μL 32P-labled oligonucleotide probe, the incubation was continued for 20 minutes on ice.
T71 1464-1629 Sentence denotes The DNA-protein complexes were separated by electrophoresis on 4% nondenaturing polyacrylamide gel in 0.5 × TBE buffer (tris-borate-EDTA) at 390 V for 1 hour at 4°C.
T2321 1464-1728 Sentence denotes The DNA-protein complexes were separated by electrophoresis on 4% nondenaturing polyacrylamide gel in 0.5 × TBE buffer (tris-borate-EDTA) at 390 V for 1 hour at 4°C. After electrophoresis, the gel was dried and exposed to X-ray film (Fuji Hyperfilm, Tokyo, Japan).
T72 1630-1728 Sentence denotes After electrophoresis, the gel was dried and exposed to X-ray film (Fuji Hyperfilm, Tokyo, Japan).
T2322 1729-1801 Sentence denotes Levels of NF-κB DNA binding activity were quantified by software ImageJ.
T73 1729-1801 Sentence denotes Levels of NF-κB DNA binding activity were quantified by software ImageJ.