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PMC:3317373 / 5505-7257 JSONTXT 9 Projects

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Id Subject Object Predicate Lexical cue
T2311 0-86 Sentence denotes Monolayers of astrocytes were washed with PBS and harvested by scraping into cold PBS.
T61 0-86 Sentence denotes Monolayers of astrocytes were washed with PBS and harvested by scraping into cold PBS.
T2312 87-279 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 87-279 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 280-381 Sentence denotes Then 10% Nonidet P-40 was added and vortexed briefly, and the nuclei were pelleted by centrifugation.
T63 280-381 Sentence denotes Then 10% Nonidet P-40 was added and vortexed briefly, and the nuclei were pelleted by centrifugation.
T2314 382-555 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 382-555 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 556-701 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 556-701 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 702-859 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 702-859 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 860-936 Sentence denotes EMSA was performed using gel shift assay system (Promega, Madison, WI, USA).
T67 860-936 Sentence denotes EMSA was performed using gel shift assay system (Promega, Madison, WI, USA).
T2318 937-1094 Sentence denotes Consensus oligonucleotide probe (5′-AGTTGAGGGGACTTTCCCAGGG-3′) was end-labeled by T4-polynucleotide kinase using [γ-32P]-ATP (Free Biotech., Beijing, China).
T68 937-1094 Sentence denotes Consensus oligonucleotide probe (5′-AGTTGAGGGGACTTTCCCAGGG-3′) was end-labeled by T4-polynucleotide kinase using [γ-32P]-ATP (Free Biotech., Beijing, China).
T2319 1095-1301 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 1095-1301 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 1302-1414 Sentence denotes After addition of the 1 μL 32P-labled oligonucleotide probe, the incubation was continued for 20 minutes on ice.
T70 1302-1414 Sentence denotes After addition of the 1 μL 32P-labled oligonucleotide probe, the incubation was continued for 20 minutes on ice.
T71 1415-1580 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 1415-1679 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 1581-1679 Sentence denotes After electrophoresis, the gel was dried and exposed to X-ray film (Fuji Hyperfilm, Tokyo, Japan).
T2322 1680-1752 Sentence denotes Levels of NF-κB DNA binding activity were quantified by software ImageJ.
T73 1680-1752 Sentence denotes Levels of NF-κB DNA binding activity were quantified by software ImageJ.