PMC:3148254 / 32170-33090 JSONTXT

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and JNK activation assays\nFor activation of CD40 signaling, 5×104 HI5 insect cells or HI5 insect cells expressing mouse CD154 (CD40 ligand) were added to 1×106 B cells. Cells were centrifuged for 1 minute at 400 x g (to promote contact between ligand cells and B cells) then incubated at 37°C for times indicated in Fig. 4. Following stimulation, cells were chilled on ice for 2 minutes. Cell pellets were dissolved in 2X SDS-PAGE loading buffer, sonicated, and heated for 5 minutes at 95°C. Total cell lysates (1×105 cell equivalents per lane) were fractionated by SDS-PAGE and blotted to polyvinylidine fluoride (PVDF) membranes. Membranes were probed with antibodies as indicated in Fig. 4. Chemiluminescent detection (Pierce Biotechnology) was used for the visualization of bands on Western blots. Images of blots were recorded with a low-light imaging system (LAS4000, Fuji Medical Systems) and on X-ray film."}

{"project":"GO-BP","denotations":[{"id":"T14231","span":{"begin":10,"end":13},"obj":"http://purl.obolibrary.org/obo/GO_0004705"},{"id":"T14232","span":{"begin":55,"end":64},"obj":"http://purl.obolibrary.org/obo/GO_0023052"}],"text":"NF-κB and JNK activation assays\nFor activation of CD40 signaling, 5×104 HI5 insect cells or HI5 insect cells expressing mouse CD154 (CD40 ligand) were added to 1×106 B cells. Cells were centrifuged for 1 minute at 400 x g (to promote contact between ligand cells and B cells) then incubated at 37°C for times indicated in Fig. 4. Following stimulation, cells were chilled on ice for 2 minutes. Cell pellets were dissolved in 2X SDS-PAGE loading buffer, sonicated, and heated for 5 minutes at 95°C. Total cell lysates (1×105 cell equivalents per lane) were fractionated by SDS-PAGE and blotted to polyvinylidine fluoride (PVDF) membranes. Membranes were probed with antibodies as indicated in Fig. 4. Chemiluminescent detection (Pierce Biotechnology) was used for the visualization of bands on Western blots. Images of blots were recorded with a low-light imaging system (LAS4000, Fuji Medical Systems) and on X-ray film."}

{"project":"GO-MF","denotations":[{"id":"T14233","span":{"begin":10,"end":13},"obj":"http://purl.obolibrary.org/obo/GO_0004705"},{"id":"T14234","span":{"begin":138,"end":144},"obj":"http://purl.obolibrary.org/obo/GO_0005488"},{"id":"T14235","span":{"begin":250,"end":256},"obj":"http://purl.obolibrary.org/obo/GO_0005488"},{"id":"T14236","span":{"begin":665,"end":675},"obj":"http://purl.obolibrary.org/obo/GO_0003823"}],"text":"NF-κB and JNK activation assays\nFor activation of CD40 signaling, 5×104 HI5 insect cells or HI5 insect cells expressing mouse CD154 (CD40 ligand) were added to 1×106 B cells. Cells were centrifuged for 1 minute at 400 x g (to promote contact between ligand cells and B cells) then incubated at 37°C for times indicated in Fig. 4. Following stimulation, cells were chilled on ice for 2 minutes. Cell pellets were dissolved in 2X SDS-PAGE loading buffer, sonicated, and heated for 5 minutes at 95°C. Total cell lysates (1×105 cell equivalents per lane) were fractionated by SDS-PAGE and blotted to polyvinylidine fluoride (PVDF) membranes. Membranes were probed with antibodies as indicated in Fig. 4. Chemiluminescent detection (Pierce Biotechnology) was used for the visualization of bands on Western blots. Images of blots were recorded with a low-light imaging system (LAS4000, Fuji Medical Systems) and on X-ray film."}

{"project":"GO-CC","denotations":[{"id":"T14242","span":{"begin":353,"end":358},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T14243","span":{"begin":394,"end":398},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T14244","span":{"begin":504,"end":508},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T14245","span":{"begin":524,"end":528},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T14246","span":{"begin":627,"end":636},"obj":"http://purl.obolibrary.org/obo/GO_0016020"},{"id":"T14247","span":{"begin":638,"end":647},"obj":"http://purl.obolibrary.org/obo/GO_0016020"},{"id":"T14248","span":{"begin":665,"end":675},"obj":"http://purl.obolibrary.org/obo/GO_0019815"},{"id":"T14249","span":{"begin":665,"end":675},"obj":"http://purl.obolibrary.org/obo/GO_0042571"},{"id":"T14237","span":{"begin":83,"end":88},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T14238","span":{"begin":103,"end":108},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T14239","span":{"begin":168,"end":173},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T14240","span":{"begin":257,"end":262},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T14241","span":{"begin":269,"end":274},"obj":"http://purl.obolibrary.org/obo/GO_0005623"}],"text":"NF-κB and JNK activation assays\nFor activation of CD40 signaling, 5×104 HI5 insect cells or HI5 insect cells expressing mouse CD154 (CD40 ligand) were added to 1×106 B cells. Cells were centrifuged for 1 minute at 400 x g (to promote contact between ligand cells and B cells) then incubated at 37°C for times indicated in Fig. 4. Following stimulation, cells were chilled on ice for 2 minutes. Cell pellets were dissolved in 2X SDS-PAGE loading buffer, sonicated, and heated for 5 minutes at 95°C. Total cell lysates (1×105 cell equivalents per lane) were fractionated by SDS-PAGE and blotted to polyvinylidine fluoride (PVDF) membranes. Membranes were probed with antibodies as indicated in Fig. 4. Chemiluminescent detection (Pierce Biotechnology) was used for the visualization of bands on Western blots. Images of blots were recorded with a low-light imaging system (LAS4000, Fuji Medical Systems) and on X-ray film."}

{"project":"sentences","denotations":[{"id":"T13868","span":{"begin":0,"end":31},"obj":"Sentence"},{"id":"T13869","span":{"begin":32,"end":174},"obj":"Sentence"},{"id":"T13870","span":{"begin":175,"end":329},"obj":"Sentence"},{"id":"T13871","span":{"begin":330,"end":393},"obj":"Sentence"},{"id":"T13872","span":{"begin":394,"end":637},"obj":"Sentence"},{"id":"T13873","span":{"begin":638,"end":699},"obj":"Sentence"},{"id":"T13874","span":{"begin":700,"end":807},"obj":"Sentence"},{"id":"T13875","span":{"begin":808,"end":920},"obj":"Sentence"},{"id":"T222","span":{"begin":0,"end":31},"obj":"Sentence"},{"id":"T223","span":{"begin":32,"end":174},"obj":"Sentence"},{"id":"T224","span":{"begin":175,"end":329},"obj":"Sentence"},{"id":"T225","span":{"begin":330,"end":393},"obj":"Sentence"},{"id":"T226","span":{"begin":394,"end":497},"obj":"Sentence"},{"id":"T227","span":{"begin":498,"end":637},"obj":"Sentence"},{"id":"T228","span":{"begin":638,"end":699},"obj":"Sentence"},{"id":"T229","span":{"begin":700,"end":807},"obj":"Sentence"},{"id":"T230","span":{"begin":808,"end":920},"obj":"Sentence"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/ontology/tao.owl#"}],"text":"NF-κB and JNK activation assays\nFor activation of CD40 signaling, 5×104 HI5 insect cells or HI5 insect cells expressing mouse CD154 (CD40 ligand) were added to 1×106 B cells. Cells were centrifuged for 1 minute at 400 x g (to promote contact between ligand cells and B cells) then incubated at 37°C for times indicated in Fig. 4. Following stimulation, cells were chilled on ice for 2 minutes. Cell pellets were dissolved in 2X SDS-PAGE loading buffer, sonicated, and heated for 5 minutes at 95°C. Total cell lysates (1×105 cell equivalents per lane) were fractionated by SDS-PAGE and blotted to polyvinylidine fluoride (PVDF) membranes. Membranes were probed with antibodies as indicated in Fig. 4. Chemiluminescent detection (Pierce Biotechnology) was used for the visualization of bands on Western blots. Images of blots were recorded with a low-light imaging system (LAS4000, Fuji Medical Systems) and on X-ray film."}

{"project":"bionlp-st-ge-2016-uniprot","denotations":[{"id":"T14219","span":{"begin":50,"end":54},"obj":"P25942"},{"id":"T14220","span":{"begin":126,"end":131},"obj":"P29965"},{"id":"T14221","span":{"begin":133,"end":137},"obj":"P25942"},{"id":"T14222","span":{"begin":133,"end":144},"obj":"P29965"}],"namespaces":[{"prefix":"_base","uri":"http://www.uniprot.org/uniprot/"}],"text":"NF-κB and JNK activation assays\nFor activation of CD40 signaling, 5×104 HI5 insect cells or HI5 insect cells expressing mouse CD154 (CD40 ligand) were added to 1×106 B cells. Cells were centrifuged for 1 minute at 400 x g (to promote contact between ligand cells and B cells) then incubated at 37°C for times indicated in Fig. 4. Following stimulation, cells were chilled on ice for 2 minutes. Cell pellets were dissolved in 2X SDS-PAGE loading buffer, sonicated, and heated for 5 minutes at 95°C. Total cell lysates (1×105 cell equivalents per lane) were fractionated by SDS-PAGE and blotted to polyvinylidine fluoride (PVDF) membranes. Membranes were probed with antibodies as indicated in Fig. 4. Chemiluminescent detection (Pierce Biotechnology) was used for the visualization of bands on Western blots. Images of blots were recorded with a low-light imaging system (LAS4000, Fuji Medical Systems) and on X-ray film."}

{"project":"test2","denotations":[{"id":"T13860","span":{"begin":50,"end":54},"obj":"Protein"},{"id":"T13861","span":{"begin":109,"end":119},"obj":"Gene_expression"},{"id":"T13862","span":{"begin":126,"end":131},"obj":"Protein"},{"id":"T13863","span":{"begin":133,"end":144},"obj":"Protein"}],"relations":[{"id":"R10896","pred":"themeOf","subj":"T13862","obj":"T13861"},{"id":"R10897","pred":"equivalentTo","subj":"T13863","obj":"T13862"}],"text":"NF-κB and JNK activation assays\nFor activation of CD40 signaling, 5×104 HI5 insect cells or HI5 insect cells expressing mouse CD154 (CD40 ligand) were added to 1×106 B cells. Cells were centrifuged for 1 minute at 400 x g (to promote contact between ligand cells and B cells) then incubated at 37°C for times indicated in Fig. 4. Following stimulation, cells were chilled on ice for 2 minutes. Cell pellets were dissolved in 2X SDS-PAGE loading buffer, sonicated, and heated for 5 minutes at 95°C. Total cell lysates (1×105 cell equivalents per lane) were fractionated by SDS-PAGE and blotted to polyvinylidine fluoride (PVDF) membranes. Membranes were probed with antibodies as indicated in Fig. 4. Chemiluminescent detection (Pierce Biotechnology) was used for the visualization of bands on Western blots. Images of blots were recorded with a low-light imaging system (LAS4000, Fuji Medical Systems) and on X-ray film."}