PMC:3333881 / 6562-8079 JSON TXT

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2_test

{"project":"2_test","denotations":[{"id":"22187158-14532125-77584438","span":{"begin":108,"end":110},"obj":"14532125"},{"id":"22187158-19782003-77584439","span":{"begin":325,"end":327},"obj":"19782003"}],"text":"Cells, transfection, treatments and luciferase assay\nHeLa, p50−/−p65−/− mouse embryonic fibroblasts (MEFs) (52) and 293T cells were cultured in Dulbecco's modified Eagle's medium; Jurkat, U937 cells and human peripheral blood mononuclear cells (PBMCs) were cultured in RPMI 1640. PBMCs were isolated as previously described (53). Media were supplemented with 10% heat-inactivated fetal calf serum and 2 mM l-glutamine (Lonza Cologne AG, Germany). HeLa, p50−/−p65−/− MEFs and 293T were transfected with DNA by using FuGENE HD (Roche Diagnostic GmbH, Mannheim, Germany), according to the manufacturer's protocol; total DNA amounts were equalized by transfection of pRc/CMV empty vector (Invitrogen, Carlsbad, CA, USA). For pulse-stimulation, HeLa cells were treated with phorbol 12-myristate 13-acetate (PMA; Sigma-Aldrich, St Louis, MO, USA) (20 ng/ml) for 5 min, or tumor necrosis factor-α (TNF-α; Sigma-Aldrich) (20 ng/ml) for 30 min, washed twice in complete culture medium and then returned to culture. For luciferase assays, pSV-β-Gal was co-transfected with pκBluc to monitor the transfection efficiency. Forty-eight-hour post-transfection, cells were lysed in lysis buffer of Dual Light Luciferase System (Tropix, Bedford, MA, USA) and the luciferase and β-galactosidase activities were evaluated by using Dual Light Luciferase System (Tropix) in a bioluminometer (Turner Biosystem, Sunnyvale, CA, USA). The ratio of firefly luciferase activity to β-galactosidase activity was expressed as relative light units."}

pmc-enju-pas

bionlp-st-ge-2016-spacy-parsed

UBERON-AE

{"project":"UBERON-AE","denotations":[{"id":"T4890","span":{"begin":220,"end":225},"obj":"http://purl.obolibrary.org/obo/UBERON_0000178"},{"id":"T4891","span":{"begin":391,"end":396},"obj":"http://purl.obolibrary.org/obo/UBERON_0001977"}],"text":"Cells, transfection, treatments and luciferase assay\nHeLa, p50−/−p65−/− mouse embryonic fibroblasts (MEFs) (52) and 293T cells were cultured in Dulbecco's modified Eagle's medium; Jurkat, U937 cells and human peripheral blood mononuclear cells (PBMCs) were cultured in RPMI 1640. PBMCs were isolated as previously described (53). Media were supplemented with 10% heat-inactivated fetal calf serum and 2 mM l-glutamine (Lonza Cologne AG, Germany). HeLa, p50−/−p65−/− MEFs and 293T were transfected with DNA by using FuGENE HD (Roche Diagnostic GmbH, Mannheim, Germany), according to the manufacturer's protocol; total DNA amounts were equalized by transfection of pRc/CMV empty vector (Invitrogen, Carlsbad, CA, USA). For pulse-stimulation, HeLa cells were treated with phorbol 12-myristate 13-acetate (PMA; Sigma-Aldrich, St Louis, MO, USA) (20 ng/ml) for 5 min, or tumor necrosis factor-α (TNF-α; Sigma-Aldrich) (20 ng/ml) for 30 min, washed twice in complete culture medium and then returned to culture. For luciferase assays, pSV-β-Gal was co-transfected with pκBluc to monitor the transfection efficiency. Forty-eight-hour post-transfection, cells were lysed in lysis buffer of Dual Light Luciferase System (Tropix, Bedford, MA, USA) and the luciferase and β-galactosidase activities were evaluated by using Dual Light Luciferase System (Tropix) in a bioluminometer (Turner Biosystem, Sunnyvale, CA, USA). The ratio of firefly luciferase activity to β-galactosidase activity was expressed as relative light units."}

GO-BP

GO-MF

{"project":"GO-MF","denotations":[{"id":"T5529","span":{"begin":707,"end":709},"obj":"http://purl.obolibrary.org/obo/GO_0033968"},{"id":"T5530","span":{"begin":1400,"end":1402},"obj":"http://purl.obolibrary.org/obo/GO_0033968"},{"id":"T5531","span":{"begin":1261,"end":1287},"obj":"http://purl.obolibrary.org/obo/GO_0004565"},{"id":"T5532","span":{"begin":1442,"end":1469},"obj":"http://purl.obolibrary.org/obo/GO_0004565"},{"id":"T5533","span":{"begin":1261,"end":1287},"obj":"http://purl.obolibrary.org/obo/GO_0004336"},{"id":"T5534","span":{"begin":1442,"end":1469},"obj":"http://purl.obolibrary.org/obo/GO_0004336"},{"id":"T5535","span":{"begin":1261,"end":1287},"obj":"http://purl.obolibrary.org/obo/GO_0033920"},{"id":"T5536","span":{"begin":1442,"end":1469},"obj":"http://purl.obolibrary.org/obo/GO_0033920"},{"id":"T5537","span":{"begin":1261,"end":1287},"obj":"http://purl.obolibrary.org/obo/GO_0033929"},{"id":"T5538","span":{"begin":1442,"end":1469},"obj":"http://purl.obolibrary.org/obo/GO_0033929"},{"id":"T5539","span":{"begin":1261,"end":1287},"obj":"http://purl.obolibrary.org/obo/GO_0033930"},{"id":"T5540","span":{"begin":1442,"end":1469},"obj":"http://purl.obolibrary.org/obo/GO_0033930"},{"id":"T5541","span":{"begin":1431,"end":1450},"obj":"http://purl.obolibrary.org/obo/GO_0045289"},{"id":"T5542","span":{"begin":1431,"end":1450},"obj":"http://purl.obolibrary.org/obo/GO_0047077"},{"id":"T5543","span":{"begin":1431,"end":1450},"obj":"http://purl.obolibrary.org/obo/GO_0047712"},{"id":"T5544","span":{"begin":1431,"end":1450},"obj":"http://purl.obolibrary.org/obo/GO_0050248"},{"id":"T5545","span":{"begin":1431,"end":1450},"obj":"http://purl.obolibrary.org/obo/GO_0050397"}],"text":"Cells, transfection, treatments and luciferase assay\nHeLa, p50−/−p65−/− mouse embryonic fibroblasts (MEFs) (52) and 293T cells were cultured in Dulbecco's modified Eagle's medium; Jurkat, U937 cells and human peripheral blood mononuclear cells (PBMCs) were cultured in RPMI 1640. PBMCs were isolated as previously described (53). Media were supplemented with 10% heat-inactivated fetal calf serum and 2 mM l-glutamine (Lonza Cologne AG, Germany). HeLa, p50−/−p65−/− MEFs and 293T were transfected with DNA by using FuGENE HD (Roche Diagnostic GmbH, Mannheim, Germany), according to the manufacturer's protocol; total DNA amounts were equalized by transfection of pRc/CMV empty vector (Invitrogen, Carlsbad, CA, USA). For pulse-stimulation, HeLa cells were treated with phorbol 12-myristate 13-acetate (PMA; Sigma-Aldrich, St Louis, MO, USA) (20 ng/ml) for 5 min, or tumor necrosis factor-α (TNF-α; Sigma-Aldrich) (20 ng/ml) for 30 min, washed twice in complete culture medium and then returned to culture. For luciferase assays, pSV-β-Gal was co-transfected with pκBluc to monitor the transfection efficiency. Forty-eight-hour post-transfection, cells were lysed in lysis buffer of Dual Light Luciferase System (Tropix, Bedford, MA, USA) and the luciferase and β-galactosidase activities were evaluated by using Dual Light Luciferase System (Tropix) in a bioluminometer (Turner Biosystem, Sunnyvale, CA, USA). The ratio of firefly luciferase activity to β-galactosidase activity was expressed as relative light units."}

GO-CC

{"project":"GO-CC","denotations":[{"id":"T5546","span":{"begin":121,"end":126},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T5547","span":{"begin":193,"end":198},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T5548","span":{"begin":238,"end":243},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T5549","span":{"begin":745,"end":750},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T5550","span":{"begin":1146,"end":1151},"obj":"http://purl.obolibrary.org/obo/GO_0005623"}],"text":"Cells, transfection, treatments and luciferase assay\nHeLa, p50−/−p65−/− mouse embryonic fibroblasts (MEFs) (52) and 293T cells were cultured in Dulbecco's modified Eagle's medium; Jurkat, U937 cells and human peripheral blood mononuclear cells (PBMCs) were cultured in RPMI 1640. PBMCs were isolated as previously described (53). Media were supplemented with 10% heat-inactivated fetal calf serum and 2 mM l-glutamine (Lonza Cologne AG, Germany). HeLa, p50−/−p65−/− MEFs and 293T were transfected with DNA by using FuGENE HD (Roche Diagnostic GmbH, Mannheim, Germany), according to the manufacturer's protocol; total DNA amounts were equalized by transfection of pRc/CMV empty vector (Invitrogen, Carlsbad, CA, USA). For pulse-stimulation, HeLa cells were treated with phorbol 12-myristate 13-acetate (PMA; Sigma-Aldrich, St Louis, MO, USA) (20 ng/ml) for 5 min, or tumor necrosis factor-α (TNF-α; Sigma-Aldrich) (20 ng/ml) for 30 min, washed twice in complete culture medium and then returned to culture. For luciferase assays, pSV-β-Gal was co-transfected with pκBluc to monitor the transfection efficiency. Forty-eight-hour post-transfection, cells were lysed in lysis buffer of Dual Light Luciferase System (Tropix, Bedford, MA, USA) and the luciferase and β-galactosidase activities were evaluated by using Dual Light Luciferase System (Tropix) in a bioluminometer (Turner Biosystem, Sunnyvale, CA, USA). The ratio of firefly luciferase activity to β-galactosidase activity was expressed as relative light units."}

sentences

{"project":"sentences","denotations":[{"id":"T4906","span":{"begin":0,"end":52},"obj":"Sentence"},{"id":"T4907","span":{"begin":53,"end":279},"obj":"Sentence"},{"id":"T4908","span":{"begin":280,"end":329},"obj":"Sentence"},{"id":"T4909","span":{"begin":330,"end":446},"obj":"Sentence"},{"id":"T4910","span":{"begin":447,"end":716},"obj":"Sentence"},{"id":"T4911","span":{"begin":717,"end":1005},"obj":"Sentence"},{"id":"T4912","span":{"begin":1006,"end":1109},"obj":"Sentence"},{"id":"T4913","span":{"begin":1110,"end":1409},"obj":"Sentence"},{"id":"T4914","span":{"begin":1410,"end":1517},"obj":"Sentence"},{"id":"T41","span":{"begin":0,"end":52},"obj":"Sentence"},{"id":"T42","span":{"begin":53,"end":279},"obj":"Sentence"},{"id":"T43","span":{"begin":280,"end":329},"obj":"Sentence"},{"id":"T44","span":{"begin":330,"end":446},"obj":"Sentence"},{"id":"T45","span":{"begin":447,"end":716},"obj":"Sentence"},{"id":"T46","span":{"begin":717,"end":1005},"obj":"Sentence"},{"id":"T47","span":{"begin":1006,"end":1109},"obj":"Sentence"},{"id":"T48","span":{"begin":1110,"end":1409},"obj":"Sentence"},{"id":"T49","span":{"begin":1410,"end":1517},"obj":"Sentence"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/ontology/tao.owl#"}],"text":"Cells, transfection, treatments and luciferase assay\nHeLa, p50−/−p65−/− mouse embryonic fibroblasts (MEFs) (52) and 293T cells were cultured in Dulbecco's modified Eagle's medium; Jurkat, U937 cells and human peripheral blood mononuclear cells (PBMCs) were cultured in RPMI 1640. PBMCs were isolated as previously described (53). Media were supplemented with 10% heat-inactivated fetal calf serum and 2 mM l-glutamine (Lonza Cologne AG, Germany). HeLa, p50−/−p65−/− MEFs and 293T were transfected with DNA by using FuGENE HD (Roche Diagnostic GmbH, Mannheim, Germany), according to the manufacturer's protocol; total DNA amounts were equalized by transfection of pRc/CMV empty vector (Invitrogen, Carlsbad, CA, USA). For pulse-stimulation, HeLa cells were treated with phorbol 12-myristate 13-acetate (PMA; Sigma-Aldrich, St Louis, MO, USA) (20 ng/ml) for 5 min, or tumor necrosis factor-α (TNF-α; Sigma-Aldrich) (20 ng/ml) for 30 min, washed twice in complete culture medium and then returned to culture. For luciferase assays, pSV-β-Gal was co-transfected with pκBluc to monitor the transfection efficiency. Forty-eight-hour post-transfection, cells were lysed in lysis buffer of Dual Light Luciferase System (Tropix, Bedford, MA, USA) and the luciferase and β-galactosidase activities were evaluated by using Dual Light Luciferase System (Tropix) in a bioluminometer (Turner Biosystem, Sunnyvale, CA, USA). The ratio of firefly luciferase activity to β-galactosidase activity was expressed as relative light units."}

events-check-again

{"project":"events-check-again","denotations":[{"id":"T5622","span":{"begin":36,"end":46},"obj":"Protein"},{"id":"T5623","span":{"begin":59,"end":62},"obj":"Protein"},{"id":"T5624","span":{"begin":65,"end":68},"obj":"Protein"},{"id":"T5625","span":{"begin":453,"end":456},"obj":"Protein"},{"id":"T5626","span":{"begin":459,"end":462},"obj":"Protein"},{"id":"T5627","span":{"begin":866,"end":889},"obj":"Protein"},{"id":"T5628","span":{"begin":891,"end":896},"obj":"Protein"},{"id":"T5629","span":{"begin":1010,"end":1020},"obj":"Protein"},{"id":"T5630","span":{"begin":1033,"end":1038},"obj":"Protein"},{"id":"T5631","span":{"begin":1193,"end":1203},"obj":"Protein"},{"id":"T5632","span":{"begin":1246,"end":1256},"obj":"Protein"},{"id":"T5633","span":{"begin":1261,"end":1276},"obj":"Protein"},{"id":"T5634","span":{"begin":1431,"end":1441},"obj":"Protein"},{"id":"T5635","span":{"begin":1454,"end":1469},"obj":"Protein"}],"relations":[{"id":"R4882","pred":"equivalentTo","subj":"T5628","obj":"T5627"}],"text":"Cells, transfection, treatments and luciferase assay\nHeLa, p50−/−p65−/− mouse embryonic fibroblasts (MEFs) (52) and 293T cells were cultured in Dulbecco's modified Eagle's medium; Jurkat, U937 cells and human peripheral blood mononuclear cells (PBMCs) were cultured in RPMI 1640. PBMCs were isolated as previously described (53). Media were supplemented with 10% heat-inactivated fetal calf serum and 2 mM l-glutamine (Lonza Cologne AG, Germany). HeLa, p50−/−p65−/− MEFs and 293T were transfected with DNA by using FuGENE HD (Roche Diagnostic GmbH, Mannheim, Germany), according to the manufacturer's protocol; total DNA amounts were equalized by transfection of pRc/CMV empty vector (Invitrogen, Carlsbad, CA, USA). For pulse-stimulation, HeLa cells were treated with phorbol 12-myristate 13-acetate (PMA; Sigma-Aldrich, St Louis, MO, USA) (20 ng/ml) for 5 min, or tumor necrosis factor-α (TNF-α; Sigma-Aldrich) (20 ng/ml) for 30 min, washed twice in complete culture medium and then returned to culture. For luciferase assays, pSV-β-Gal was co-transfected with pκBluc to monitor the transfection efficiency. Forty-eight-hour post-transfection, cells were lysed in lysis buffer of Dual Light Luciferase System (Tropix, Bedford, MA, USA) and the luciferase and β-galactosidase activities were evaluated by using Dual Light Luciferase System (Tropix) in a bioluminometer (Turner Biosystem, Sunnyvale, CA, USA). The ratio of firefly luciferase activity to β-galactosidase activity was expressed as relative light units."}

bionlp-st-ge-2016-reference-tees

{"project":"bionlp-st-ge-2016-reference-tees","denotations":[{"id":"T5636","span":{"begin":36,"end":46},"obj":"Protein"},{"id":"T5637","span":{"begin":515,"end":524},"obj":"Protein"},{"id":"T5638","span":{"begin":866,"end":889},"obj":"Protein"},{"id":"T5639","span":{"begin":891,"end":896},"obj":"Protein"},{"id":"T5640","span":{"begin":898,"end":911},"obj":"Protein"},{"id":"T5641","span":{"begin":1010,"end":1020},"obj":"Protein"},{"id":"T5642","span":{"begin":1063,"end":1069},"obj":"Protein"},{"id":"T5643","span":{"begin":1043,"end":1057},"obj":"Gene_expression"},{"id":"T5644","span":{"begin":1043,"end":1057},"obj":"Gene_expression"},{"id":"T5645","span":{"begin":1043,"end":1057},"obj":"Positive_regulation"},{"id":"T5646","span":{"begin":1043,"end":1057},"obj":"Positive_regulation"},{"id":"T5647","span":{"begin":1187,"end":1203},"obj":"Protein"},{"id":"T5648","span":{"begin":1246,"end":1256},"obj":"Protein"},{"id":"T5649","span":{"begin":1261,"end":1276},"obj":"Protein"},{"id":"T5650","span":{"begin":1317,"end":1333},"obj":"Protein"},{"id":"T5651","span":{"begin":1431,"end":1441},"obj":"Protein"},{"id":"T5652","span":{"begin":1454,"end":1469},"obj":"Protein"}],"relations":[{"id":"R4883","pred":"themeOf","subj":"T5641","obj":"T5643"},{"id":"R4884","pred":"themeOf","subj":"T5642","obj":"T5644"},{"id":"R4885","pred":"themeOf","subj":"T5643","obj":"T5645"},{"id":"R4886","pred":"themeOf","subj":"T5644","obj":"T5646"}],"text":"Cells, transfection, treatments and luciferase assay\nHeLa, p50−/−p65−/− mouse embryonic fibroblasts (MEFs) (52) and 293T cells were cultured in Dulbecco's modified Eagle's medium; Jurkat, U937 cells and human peripheral blood mononuclear cells (PBMCs) were cultured in RPMI 1640. PBMCs were isolated as previously described (53). Media were supplemented with 10% heat-inactivated fetal calf serum and 2 mM l-glutamine (Lonza Cologne AG, Germany). HeLa, p50−/−p65−/− MEFs and 293T were transfected with DNA by using FuGENE HD (Roche Diagnostic GmbH, Mannheim, Germany), according to the manufacturer's protocol; total DNA amounts were equalized by transfection of pRc/CMV empty vector (Invitrogen, Carlsbad, CA, USA). For pulse-stimulation, HeLa cells were treated with phorbol 12-myristate 13-acetate (PMA; Sigma-Aldrich, St Louis, MO, USA) (20 ng/ml) for 5 min, or tumor necrosis factor-α (TNF-α; Sigma-Aldrich) (20 ng/ml) for 30 min, washed twice in complete culture medium and then returned to culture. For luciferase assays, pSV-β-Gal was co-transfected with pκBluc to monitor the transfection efficiency. Forty-eight-hour post-transfection, cells were lysed in lysis buffer of Dual Light Luciferase System (Tropix, Bedford, MA, USA) and the luciferase and β-galactosidase activities were evaluated by using Dual Light Luciferase System (Tropix) in a bioluminometer (Turner Biosystem, Sunnyvale, CA, USA). The ratio of firefly luciferase activity to β-galactosidase activity was expressed as relative light units."}

test2

{"project":"test2","denotations":[{"id":"T4875","span":{"begin":36,"end":46},"obj":"Protein"},{"id":"T4876","span":{"begin":59,"end":62},"obj":"Protein"},{"id":"T4877","span":{"begin":65,"end":68},"obj":"Protein"},{"id":"T4878","span":{"begin":453,"end":456},"obj":"Protein"},{"id":"T4879","span":{"begin":459,"end":462},"obj":"Protein"},{"id":"T4880","span":{"begin":866,"end":889},"obj":"Protein"},{"id":"T4881","span":{"begin":891,"end":896},"obj":"Protein"},{"id":"T4882","span":{"begin":1010,"end":1020},"obj":"Protein"},{"id":"T4883","span":{"begin":1033,"end":1038},"obj":"Protein"},{"id":"T4884","span":{"begin":1193,"end":1203},"obj":"Protein"},{"id":"T4885","span":{"begin":1246,"end":1256},"obj":"Protein"},{"id":"T4886","span":{"begin":1261,"end":1276},"obj":"Protein"},{"id":"T4887","span":{"begin":1431,"end":1441},"obj":"Protein"},{"id":"T4888","span":{"begin":1454,"end":1469},"obj":"Protein"},{"id":"T4889","span":{"begin":1483,"end":1492},"obj":"Gene_expression"}],"relations":[{"id":"R4306","pred":"equivalentTo","subj":"T4881","obj":"T4880"}],"text":"Cells, transfection, treatments and luciferase assay\nHeLa, p50−/−p65−/− mouse embryonic fibroblasts (MEFs) (52) and 293T cells were cultured in Dulbecco's modified Eagle's medium; Jurkat, U937 cells and human peripheral blood mononuclear cells (PBMCs) were cultured in RPMI 1640. PBMCs were isolated as previously described (53). Media were supplemented with 10% heat-inactivated fetal calf serum and 2 mM l-glutamine (Lonza Cologne AG, Germany). HeLa, p50−/−p65−/− MEFs and 293T were transfected with DNA by using FuGENE HD (Roche Diagnostic GmbH, Mannheim, Germany), according to the manufacturer's protocol; total DNA amounts were equalized by transfection of pRc/CMV empty vector (Invitrogen, Carlsbad, CA, USA). For pulse-stimulation, HeLa cells were treated with phorbol 12-myristate 13-acetate (PMA; Sigma-Aldrich, St Louis, MO, USA) (20 ng/ml) for 5 min, or tumor necrosis factor-α (TNF-α; Sigma-Aldrich) (20 ng/ml) for 30 min, washed twice in complete culture medium and then returned to culture. For luciferase assays, pSV-β-Gal was co-transfected with pκBluc to monitor the transfection efficiency. Forty-eight-hour post-transfection, cells were lysed in lysis buffer of Dual Light Luciferase System (Tropix, Bedford, MA, USA) and the luciferase and β-galactosidase activities were evaluated by using Dual Light Luciferase System (Tropix) in a bioluminometer (Turner Biosystem, Sunnyvale, CA, USA). The ratio of firefly luciferase activity to β-galactosidase activity was expressed as relative light units."}

bionlp-st-ge-2016-reference

{"project":"bionlp-st-ge-2016-reference","denotations":[{"id":"T4892","span":{"begin":36,"end":46},"obj":"Protein"},{"id":"T4893","span":{"begin":59,"end":62},"obj":"Protein"},{"id":"T4894","span":{"begin":65,"end":68},"obj":"Protein"},{"id":"T4895","span":{"begin":453,"end":456},"obj":"Protein"},{"id":"T4896","span":{"begin":459,"end":462},"obj":"Protein"},{"id":"T4897","span":{"begin":866,"end":889},"obj":"Protein"},{"id":"T4898","span":{"begin":891,"end":896},"obj":"Protein"},{"id":"T4899","span":{"begin":1010,"end":1020},"obj":"Protein"},{"id":"T4900","span":{"begin":1033,"end":1038},"obj":"Protein"},{"id":"T4901","span":{"begin":1193,"end":1203},"obj":"Protein"},{"id":"T4902","span":{"begin":1246,"end":1256},"obj":"Protein"},{"id":"T4903","span":{"begin":1261,"end":1276},"obj":"Protein"},{"id":"T4904","span":{"begin":1431,"end":1441},"obj":"Protein"},{"id":"T4905","span":{"begin":1454,"end":1469},"obj":"Protein"}],"relations":[{"id":"R4307","pred":"equivalentTo","subj":"T4898","obj":"T4897"}],"namespaces":[{"prefix":"_base","uri":"http://bionlp.dbcls.jp/ontology/ge.owl#"}],"text":"Cells, transfection, treatments and luciferase assay\nHeLa, p50−/−p65−/− mouse embryonic fibroblasts (MEFs) (52) and 293T cells were cultured in Dulbecco's modified Eagle's medium; Jurkat, U937 cells and human peripheral blood mononuclear cells (PBMCs) were cultured in RPMI 1640. PBMCs were isolated as previously described (53). Media were supplemented with 10% heat-inactivated fetal calf serum and 2 mM l-glutamine (Lonza Cologne AG, Germany). HeLa, p50−/−p65−/− MEFs and 293T were transfected with DNA by using FuGENE HD (Roche Diagnostic GmbH, Mannheim, Germany), according to the manufacturer's protocol; total DNA amounts were equalized by transfection of pRc/CMV empty vector (Invitrogen, Carlsbad, CA, USA). For pulse-stimulation, HeLa cells were treated with phorbol 12-myristate 13-acetate (PMA; Sigma-Aldrich, St Louis, MO, USA) (20 ng/ml) for 5 min, or tumor necrosis factor-α (TNF-α; Sigma-Aldrich) (20 ng/ml) for 30 min, washed twice in complete culture medium and then returned to culture. For luciferase assays, pSV-β-Gal was co-transfected with pκBluc to monitor the transfection efficiency. Forty-eight-hour post-transfection, cells were lysed in lysis buffer of Dual Light Luciferase System (Tropix, Bedford, MA, USA) and the luciferase and β-galactosidase activities were evaluated by using Dual Light Luciferase System (Tropix) in a bioluminometer (Turner Biosystem, Sunnyvale, CA, USA). The ratio of firefly luciferase activity to β-galactosidase activity was expressed as relative light units."}

bionlp-st-ge-2016-uniprot

{"project":"bionlp-st-ge-2016-uniprot","denotations":[{"id":"T5183","span":{"begin":36,"end":46},"obj":"P08659"},{"id":"T5184","span":{"begin":59,"end":62},"obj":"P19838"},{"id":"T5185","span":{"begin":65,"end":68},"obj":"Q04206"},{"id":"T5186","span":{"begin":65,"end":68},"obj":"P21579"},{"id":"T5187","span":{"begin":453,"end":456},"obj":"P19838"},{"id":"T5188","span":{"begin":459,"end":462},"obj":"P21579"},{"id":"T5189","span":{"begin":459,"end":462},"obj":"Q04206"},{"id":"T5190","span":{"begin":866,"end":887},"obj":"P01375"},{"id":"T5191","span":{"begin":891,"end":894},"obj":"P01375"},{"id":"T5192","span":{"begin":891,"end":896},"obj":"P01375"},{"id":"T5193","span":{"begin":1010,"end":1020},"obj":"P08659"},{"id":"T5194","span":{"begin":1193,"end":1203},"obj":"P08659"},{"id":"T5195","span":{"begin":1246,"end":1256},"obj":"P08659"},{"id":"T5196","span":{"begin":1323,"end":1333},"obj":"P08659"},{"id":"T5197","span":{"begin":1431,"end":1441},"obj":"P08659"},{"id":"T5198","span":{"begin":1496,"end":1504},"obj":"Q04864"}],"namespaces":[{"prefix":"_base","uri":"http://www.uniprot.org/uniprot/"}],"text":"Cells, transfection, treatments and luciferase assay\nHeLa, p50−/−p65−/− mouse embryonic fibroblasts (MEFs) (52) and 293T cells were cultured in Dulbecco's modified Eagle's medium; Jurkat, U937 cells and human peripheral blood mononuclear cells (PBMCs) were cultured in RPMI 1640. PBMCs were isolated as previously described (53). Media were supplemented with 10% heat-inactivated fetal calf serum and 2 mM l-glutamine (Lonza Cologne AG, Germany). HeLa, p50−/−p65−/− MEFs and 293T were transfected with DNA by using FuGENE HD (Roche Diagnostic GmbH, Mannheim, Germany), according to the manufacturer's protocol; total DNA amounts were equalized by transfection of pRc/CMV empty vector (Invitrogen, Carlsbad, CA, USA). For pulse-stimulation, HeLa cells were treated with phorbol 12-myristate 13-acetate (PMA; Sigma-Aldrich, St Louis, MO, USA) (20 ng/ml) for 5 min, or tumor necrosis factor-α (TNF-α; Sigma-Aldrich) (20 ng/ml) for 30 min, washed twice in complete culture medium and then returned to culture. For luciferase assays, pSV-β-Gal was co-transfected with pκBluc to monitor the transfection efficiency. Forty-eight-hour post-transfection, cells were lysed in lysis buffer of Dual Light Luciferase System (Tropix, Bedford, MA, USA) and the luciferase and β-galactosidase activities were evaluated by using Dual Light Luciferase System (Tropix) in a bioluminometer (Turner Biosystem, Sunnyvale, CA, USA). The ratio of firefly luciferase activity to β-galactosidase activity was expressed as relative light units."}

PMC:3333881 / 6562-8079 JSONTXT

Annnotations TAB JSON ListView MergeView

PMC:3333881 / 6562-8079 JSON TXT