PMC:1064895 / 10999-12494 JSONTXT

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After 16 hours of incubation, mRNA was extracted with RNAzol B (Biotex Laboratories, Houston, TX, USA) in accordance with the manufacturer's instructions. Reverse transcription of 2 μg of total mRNA was performed at 42°C using the Superscript™ reverse transcription system (Takara, Shiga, Japan). PCR amplification of cDNA aliquots was performed by adding 2.5 mM dNTPs, 2.5 U of Taq DNA polymerase (Takara) and 0.25 μM of sense and antisense primers. The reaction was performed in PCR buffer (1.5 mM MgCl2, 50 mM KCl, 10 mM Tris-HCl, pH 8.3) in a total volume of 25 μl. The following sense and antisense primers for each molecules were used: IL-17 sense, 5'-ATG ACT CCT GGG AAG ACC TCA TTG-3'; IL-17 antisense, 5'-TTA GGC CAC ATG GTG GAC AAT CGG-3'; glyceraldehyde-3-phosphate dehydrogenase (GAPDH) sense, 5'-CGA TGC TGG GCG TGA GTA C-3'; GAPDH antisense, 5'-CGT TCA GCT CAG GGA TGA CC-3'. Reactions were processed in a DNA thermal cycler (Perkin-Elmer Cetus, Norwalk, CT, USA) through cycles for 30 s of denaturation at 94°C, 1 min of annealing at 56°C for GAPDH and IL-17, followed by 1 min of elongation at 72°C. PCR rounds were repeated for 25 cycles each for both GAPDH and IL-17; this was determined as falling within the exponential phase of amplification for each molecule. The level of mRNA expression was presented as a ratio of IL-17 PCR product over GAPDH product."}

{"project":"bionlp-st-ge-2016-test-proteins","denotations":[{"id":"T8664","span":{"begin":1481,"end":1486},"obj":"Protein"},{"id":"T8663","span":{"begin":1458,"end":1463},"obj":"Protein"},{"id":"T8662","span":{"begin":1298,"end":1303},"obj":"Protein"},{"id":"T8661","span":{"begin":1288,"end":1293},"obj":"Protein"},{"id":"T8660","span":{"begin":1187,"end":1192},"obj":"Protein"},{"id":"T8659","span":{"begin":1177,"end":1182},"obj":"Protein"},{"id":"T8658","span":{"begin":958,"end":963},"obj":"Protein"},{"id":"T8657","span":{"begin":911,"end":916},"obj":"Protein"},{"id":"T8656","span":{"begin":869,"end":909},"obj":"Protein"},{"id":"T8655","span":{"begin":761,"end":766},"obj":"Protein"},{"id":"T8654","span":{"begin":498,"end":501},"obj":"Protein"}],"namespaces":[{"prefix":"_base","uri":"http://bionlp.dbcls.jp/ontology/ge.owl#"}],"text":"PBMC were incubated with various concentrations of anti-CD3 in the presence or absence of inhibitors (LY294002, PDTC). After 16 hours of incubation, mRNA was extracted with RNAzol B (Biotex Laboratories, Houston, TX, USA) in accordance with the manufacturer's instructions. Reverse transcription of 2 μg of total mRNA was performed at 42°C using the Superscript™ reverse transcription system (Takara, Shiga, Japan). PCR amplification of cDNA aliquots was performed by adding 2.5 mM dNTPs, 2.5 U of Taq DNA polymerase (Takara) and 0.25 μM of sense and antisense primers. The reaction was performed in PCR buffer (1.5 mM MgCl2, 50 mM KCl, 10 mM Tris-HCl, pH 8.3) in a total volume of 25 μl. The following sense and antisense primers for each molecules were used: IL-17 sense, 5'-ATG ACT CCT GGG AAG ACC TCA TTG-3'; IL-17 antisense, 5'-TTA GGC CAC ATG GTG GAC AAT CGG-3'; glyceraldehyde-3-phosphate dehydrogenase (GAPDH) sense, 5'-CGA TGC TGG GCG TGA GTA C-3'; GAPDH antisense, 5'-CGT TCA GCT CAG GGA TGA CC-3'. Reactions were processed in a DNA thermal cycler (Perkin-Elmer Cetus, Norwalk, CT, USA) through cycles for 30 s of denaturation at 94°C, 1 min of annealing at 56°C for GAPDH and IL-17, followed by 1 min of elongation at 72°C. PCR rounds were repeated for 25 cycles each for both GAPDH and IL-17; this was determined as falling within the exponential phase of amplification for each molecule. The level of mRNA expression was presented as a ratio of IL-17 PCR product over GAPDH product."}

{"project":"bionlp-st-ge-2016-uniprot","denotations":[{"id":"T8726","span":{"begin":932,"end":935},"obj":"http://www.uniprot.org/uniprot/P21980"},{"id":"T8725","span":{"begin":1481,"end":1486},"obj":"http://www.uniprot.org/uniprot/P04406"},{"id":"T8724","span":{"begin":1288,"end":1293},"obj":"http://www.uniprot.org/uniprot/P04406"},{"id":"T8723","span":{"begin":1177,"end":1182},"obj":"http://www.uniprot.org/uniprot/P04406"},{"id":"T8722","span":{"begin":958,"end":963},"obj":"http://www.uniprot.org/uniprot/P04406"},{"id":"T8721","span":{"begin":911,"end":916},"obj":"http://www.uniprot.org/uniprot/P04406"},{"id":"T8720","span":{"begin":884,"end":909},"obj":"http://www.uniprot.org/uniprot/P04406"},{"id":"T8719","span":{"begin":56,"end":59},"obj":"http://www.uniprot.org/uniprot/P09693"},{"id":"T8718","span":{"begin":56,"end":59},"obj":"http://www.uniprot.org/uniprot/P07766"},{"id":"T8717","span":{"begin":56,"end":59},"obj":"http://www.uniprot.org/uniprot/P04234"},{"id":"T8716","span":{"begin":56,"end":59},"obj":"http://www.uniprot.org/uniprot/P20963"},{"id":"T8715","span":{"begin":1458,"end":1463},"obj":"http://www.uniprot.org/uniprot/Q16552"},{"id":"T8714","span":{"begin":1298,"end":1303},"obj":"http://www.uniprot.org/uniprot/Q16552"},{"id":"T8713","span":{"begin":1187,"end":1192},"obj":"http://www.uniprot.org/uniprot/Q16552"},{"id":"T8712","span":{"begin":813,"end":818},"obj":"http://www.uniprot.org/uniprot/Q16552"},{"id":"T8711","span":{"begin":761,"end":766},"obj":"http://www.uniprot.org/uniprot/Q16552"}],"namespaces":[{"prefix":"_base","uri":"http://www.uniprot.org/uniprot/"}],"text":"PBMC were incubated with various concentrations of anti-CD3 in the presence or absence of inhibitors (LY294002, PDTC). After 16 hours of incubation, mRNA was extracted with RNAzol B (Biotex Laboratories, Houston, TX, USA) in accordance with the manufacturer's instructions. Reverse transcription of 2 μg of total mRNA was performed at 42°C using the Superscript™ reverse transcription system (Takara, Shiga, Japan). PCR amplification of cDNA aliquots was performed by adding 2.5 mM dNTPs, 2.5 U of Taq DNA polymerase (Takara) and 0.25 μM of sense and antisense primers. The reaction was performed in PCR buffer (1.5 mM MgCl2, 50 mM KCl, 10 mM Tris-HCl, pH 8.3) in a total volume of 25 μl. The following sense and antisense primers for each molecules were used: IL-17 sense, 5'-ATG ACT CCT GGG AAG ACC TCA TTG-3'; IL-17 antisense, 5'-TTA GGC CAC ATG GTG GAC AAT CGG-3'; glyceraldehyde-3-phosphate dehydrogenase (GAPDH) sense, 5'-CGA TGC TGG GCG TGA GTA C-3'; GAPDH antisense, 5'-CGT TCA GCT CAG GGA TGA CC-3'. Reactions were processed in a DNA thermal cycler (Perkin-Elmer Cetus, Norwalk, CT, USA) through cycles for 30 s of denaturation at 94°C, 1 min of annealing at 56°C for GAPDH and IL-17, followed by 1 min of elongation at 72°C. PCR rounds were repeated for 25 cycles each for both GAPDH and IL-17; this was determined as falling within the exponential phase of amplification for each molecule. The level of mRNA expression was presented as a ratio of IL-17 PCR product over GAPDH product."}

{"project":"GO-BP","denotations":[{"id":"T8638","span":{"begin":978,"end":981},"obj":"http://purl.obolibrary.org/obo/GO_0047801"},{"id":"T8637","span":{"begin":857,"end":860},"obj":"http://purl.obolibrary.org/obo/GO_0004069"},{"id":"T8636","span":{"begin":371,"end":384},"obj":"http://purl.obolibrary.org/obo/GO_0006351"},{"id":"T8635","span":{"begin":282,"end":295},"obj":"http://purl.obolibrary.org/obo/GO_0006351"},{"id":"T8633","span":{"begin":363,"end":384},"obj":"http://purl.obolibrary.org/obo/GO_0001171"}],"text":"PBMC were incubated with various concentrations of anti-CD3 in the presence or absence of inhibitors (LY294002, PDTC). After 16 hours of incubation, mRNA was extracted with RNAzol B (Biotex Laboratories, Houston, TX, USA) in accordance with the manufacturer's instructions. Reverse transcription of 2 μg of total mRNA was performed at 42°C using the Superscript™ reverse transcription system (Takara, Shiga, Japan). PCR amplification of cDNA aliquots was performed by adding 2.5 mM dNTPs, 2.5 U of Taq DNA polymerase (Takara) and 0.25 μM of sense and antisense primers. The reaction was performed in PCR buffer (1.5 mM MgCl2, 50 mM KCl, 10 mM Tris-HCl, pH 8.3) in a total volume of 25 μl. The following sense and antisense primers for each molecules were used: IL-17 sense, 5'-ATG ACT CCT GGG AAG ACC TCA TTG-3'; IL-17 antisense, 5'-TTA GGC CAC ATG GTG GAC AAT CGG-3'; glyceraldehyde-3-phosphate dehydrogenase (GAPDH) sense, 5'-CGA TGC TGG GCG TGA GTA C-3'; GAPDH antisense, 5'-CGT TCA GCT CAG GGA TGA CC-3'. Reactions were processed in a DNA thermal cycler (Perkin-Elmer Cetus, Norwalk, CT, USA) through cycles for 30 s of denaturation at 94°C, 1 min of annealing at 56°C for GAPDH and IL-17, followed by 1 min of elongation at 72°C. PCR rounds were repeated for 25 cycles each for both GAPDH and IL-17; this was determined as falling within the exponential phase of amplification for each molecule. The level of mRNA expression was presented as a ratio of IL-17 PCR product over GAPDH product."}

{"project":"GO-MF","denotations":[{"id":"T8673","span":{"begin":978,"end":981},"obj":"http://purl.obolibrary.org/obo/GO_0047801"},{"id":"T8672","span":{"begin":857,"end":860},"obj":"http://purl.obolibrary.org/obo/GO_0004069"},{"id":"T8671","span":{"begin":809,"end":815},"obj":"http://purl.obolibrary.org/obo/GO_0005135"},{"id":"T8670","span":{"begin":1458,"end":1463},"obj":"http://purl.obolibrary.org/obo/GO_0030367"},{"id":"T8669","span":{"begin":1298,"end":1303},"obj":"http://purl.obolibrary.org/obo/GO_0030367"},{"id":"T8668","span":{"begin":1187,"end":1192},"obj":"http://purl.obolibrary.org/obo/GO_0030367"},{"id":"T8667","span":{"begin":813,"end":818},"obj":"http://purl.obolibrary.org/obo/GO_0030367"},{"id":"T8666","span":{"begin":761,"end":766},"obj":"http://purl.obolibrary.org/obo/GO_0030367"}],"text":"PBMC were incubated with various concentrations of anti-CD3 in the presence or absence of inhibitors (LY294002, PDTC). After 16 hours of incubation, mRNA was extracted with RNAzol B (Biotex Laboratories, Houston, TX, USA) in accordance with the manufacturer's instructions. Reverse transcription of 2 μg of total mRNA was performed at 42°C using the Superscript™ reverse transcription system (Takara, Shiga, Japan). PCR amplification of cDNA aliquots was performed by adding 2.5 mM dNTPs, 2.5 U of Taq DNA polymerase (Takara) and 0.25 μM of sense and antisense primers. The reaction was performed in PCR buffer (1.5 mM MgCl2, 50 mM KCl, 10 mM Tris-HCl, pH 8.3) in a total volume of 25 μl. The following sense and antisense primers for each molecules were used: IL-17 sense, 5'-ATG ACT CCT GGG AAG ACC TCA TTG-3'; IL-17 antisense, 5'-TTA GGC CAC ATG GTG GAC AAT CGG-3'; glyceraldehyde-3-phosphate dehydrogenase (GAPDH) sense, 5'-CGA TGC TGG GCG TGA GTA C-3'; GAPDH antisense, 5'-CGT TCA GCT CAG GGA TGA CC-3'. Reactions were processed in a DNA thermal cycler (Perkin-Elmer Cetus, Norwalk, CT, USA) through cycles for 30 s of denaturation at 94°C, 1 min of annealing at 56°C for GAPDH and IL-17, followed by 1 min of elongation at 72°C. PCR rounds were repeated for 25 cycles each for both GAPDH and IL-17; this was determined as falling within the exponential phase of amplification for each molecule. The level of mRNA expression was presented as a ratio of IL-17 PCR product over GAPDH product."}

{"project":"sentences","denotations":[{"id":"T8631","span":{"begin":1401,"end":1495},"obj":"Sentence"},{"id":"T8630","span":{"begin":1009,"end":1400},"obj":"Sentence"},{"id":"T8629","span":{"begin":689,"end":1008},"obj":"Sentence"},{"id":"T8628","span":{"begin":570,"end":688},"obj":"Sentence"},{"id":"T8627","span":{"begin":416,"end":569},"obj":"Sentence"},{"id":"T8626","span":{"begin":274,"end":415},"obj":"Sentence"},{"id":"T8625","span":{"begin":119,"end":273},"obj":"Sentence"},{"id":"T8624","span":{"begin":0,"end":118},"obj":"Sentence"},{"id":"T83","span":{"begin":0,"end":118},"obj":"Sentence"},{"id":"T84","span":{"begin":119,"end":273},"obj":"Sentence"},{"id":"T85","span":{"begin":274,"end":415},"obj":"Sentence"},{"id":"T86","span":{"begin":416,"end":569},"obj":"Sentence"},{"id":"T87","span":{"begin":570,"end":688},"obj":"Sentence"},{"id":"T88","span":{"begin":689,"end":1008},"obj":"Sentence"},{"id":"T89","span":{"begin":1009,"end":1234},"obj":"Sentence"},{"id":"T90","span":{"begin":1235,"end":1400},"obj":"Sentence"},{"id":"T91","span":{"begin":1401,"end":1495},"obj":"Sentence"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/ontology/tao.owl#"}],"text":"PBMC were incubated with various concentrations of anti-CD3 in the presence or absence of inhibitors (LY294002, PDTC). After 16 hours of incubation, mRNA was extracted with RNAzol B (Biotex Laboratories, Houston, TX, USA) in accordance with the manufacturer's instructions. Reverse transcription of 2 μg of total mRNA was performed at 42°C using the Superscript™ reverse transcription system (Takara, Shiga, Japan). PCR amplification of cDNA aliquots was performed by adding 2.5 mM dNTPs, 2.5 U of Taq DNA polymerase (Takara) and 0.25 μM of sense and antisense primers. The reaction was performed in PCR buffer (1.5 mM MgCl2, 50 mM KCl, 10 mM Tris-HCl, pH 8.3) in a total volume of 25 μl. The following sense and antisense primers for each molecules were used: IL-17 sense, 5'-ATG ACT CCT GGG AAG ACC TCA TTG-3'; IL-17 antisense, 5'-TTA GGC CAC ATG GTG GAC AAT CGG-3'; glyceraldehyde-3-phosphate dehydrogenase (GAPDH) sense, 5'-CGA TGC TGG GCG TGA GTA C-3'; GAPDH antisense, 5'-CGT TCA GCT CAG GGA TGA CC-3'. Reactions were processed in a DNA thermal cycler (Perkin-Elmer Cetus, Norwalk, CT, USA) through cycles for 30 s of denaturation at 94°C, 1 min of annealing at 56°C for GAPDH and IL-17, followed by 1 min of elongation at 72°C. PCR rounds were repeated for 25 cycles each for both GAPDH and IL-17; this was determined as falling within the exponential phase of amplification for each molecule. The level of mRNA expression was presented as a ratio of IL-17 PCR product over GAPDH product."}

{"project":"simple1","denotations":[{"id":"T8709","span":{"begin":1481,"end":1486},"obj":"Protein"},{"id":"T8708","span":{"begin":1458,"end":1463},"obj":"Protein"},{"id":"T8707","span":{"begin":1298,"end":1303},"obj":"Protein"},{"id":"T8706","span":{"begin":1288,"end":1293},"obj":"Protein"},{"id":"T8705","span":{"begin":1187,"end":1192},"obj":"Protein"},{"id":"T8704","span":{"begin":1177,"end":1182},"obj":"Protein"},{"id":"T8703","span":{"begin":958,"end":963},"obj":"Protein"},{"id":"T8702","span":{"begin":911,"end":916},"obj":"Protein"},{"id":"T8701","span":{"begin":869,"end":909},"obj":"Protein"},{"id":"T8700","span":{"begin":761,"end":766},"obj":"Protein"},{"id":"T8699","span":{"begin":498,"end":501},"obj":"Protein"}],"text":"PBMC were incubated with various concentrations of anti-CD3 in the presence or absence of inhibitors (LY294002, PDTC). After 16 hours of incubation, mRNA was extracted with RNAzol B (Biotex Laboratories, Houston, TX, USA) in accordance with the manufacturer's instructions. Reverse transcription of 2 μg of total mRNA was performed at 42°C using the Superscript™ reverse transcription system (Takara, Shiga, Japan). PCR amplification of cDNA aliquots was performed by adding 2.5 mM dNTPs, 2.5 U of Taq DNA polymerase (Takara) and 0.25 μM of sense and antisense primers. The reaction was performed in PCR buffer (1.5 mM MgCl2, 50 mM KCl, 10 mM Tris-HCl, pH 8.3) in a total volume of 25 μl. The following sense and antisense primers for each molecules were used: IL-17 sense, 5'-ATG ACT CCT GGG AAG ACC TCA TTG-3'; IL-17 antisense, 5'-TTA GGC CAC ATG GTG GAC AAT CGG-3'; glyceraldehyde-3-phosphate dehydrogenase (GAPDH) sense, 5'-CGA TGC TGG GCG TGA GTA C-3'; GAPDH antisense, 5'-CGT TCA GCT CAG GGA TGA CC-3'. Reactions were processed in a DNA thermal cycler (Perkin-Elmer Cetus, Norwalk, CT, USA) through cycles for 30 s of denaturation at 94°C, 1 min of annealing at 56°C for GAPDH and IL-17, followed by 1 min of elongation at 72°C. PCR rounds were repeated for 25 cycles each for both GAPDH and IL-17; this was determined as falling within the exponential phase of amplification for each molecule. The level of mRNA expression was presented as a ratio of IL-17 PCR product over GAPDH product."}

{"project":"DLUT931","denotations":[{"id":"T8753","span":{"begin":1487,"end":1494},"obj":"Gene_expression"},{"id":"T8752","span":{"begin":964,"end":973},"obj":"Negative_regulation"},{"id":"T8750","span":{"begin":1481,"end":1486},"obj":"Protein"},{"id":"T8749","span":{"begin":1458,"end":1463},"obj":"Protein"},{"id":"T8748","span":{"begin":1288,"end":1293},"obj":"Protein"},{"id":"T8747","span":{"begin":1298,"end":1303},"obj":"Protein"},{"id":"T8746","span":{"begin":1187,"end":1192},"obj":"Protein"},{"id":"T8745","span":{"begin":1177,"end":1182},"obj":"Protein"},{"id":"T8744","span":{"begin":958,"end":963},"obj":"Protein"},{"id":"T8743","span":{"begin":911,"end":916},"obj":"Protein"},{"id":"T8742","span":{"begin":869,"end":909},"obj":"Protein"},{"id":"T8741","span":{"begin":761,"end":766},"obj":"Protein"},{"id":"T8740","span":{"begin":498,"end":501},"obj":"Protein"}],"relations":[{"id":"R4483","pred":"themeOf","subj":"T8741","obj":"T8752"},{"id":"R4484","pred":"themeOf","subj":"T8744","obj":"T8752"},{"id":"R4485","pred":"themeOf","subj":"T8750","obj":"T8753"}],"text":"PBMC were incubated with various concentrations of anti-CD3 in the presence or absence of inhibitors (LY294002, PDTC). After 16 hours of incubation, mRNA was extracted with RNAzol B (Biotex Laboratories, Houston, TX, USA) in accordance with the manufacturer's instructions. Reverse transcription of 2 μg of total mRNA was performed at 42°C using the Superscript™ reverse transcription system (Takara, Shiga, Japan). PCR amplification of cDNA aliquots was performed by adding 2.5 mM dNTPs, 2.5 U of Taq DNA polymerase (Takara) and 0.25 μM of sense and antisense primers. The reaction was performed in PCR buffer (1.5 mM MgCl2, 50 mM KCl, 10 mM Tris-HCl, pH 8.3) in a total volume of 25 μl. The following sense and antisense primers for each molecules were used: IL-17 sense, 5'-ATG ACT CCT GGG AAG ACC TCA TTG-3'; IL-17 antisense, 5'-TTA GGC CAC ATG GTG GAC AAT CGG-3'; glyceraldehyde-3-phosphate dehydrogenase (GAPDH) sense, 5'-CGA TGC TGG GCG TGA GTA C-3'; GAPDH antisense, 5'-CGT TCA GCT CAG GGA TGA CC-3'. Reactions were processed in a DNA thermal cycler (Perkin-Elmer Cetus, Norwalk, CT, USA) through cycles for 30 s of denaturation at 94°C, 1 min of annealing at 56°C for GAPDH and IL-17, followed by 1 min of elongation at 72°C. PCR rounds were repeated for 25 cycles each for both GAPDH and IL-17; this was determined as falling within the exponential phase of amplification for each molecule. The level of mRNA expression was presented as a ratio of IL-17 PCR product over GAPDH product."}

{"project":"bionlp-st-ge-2016-test-ihmc","denotations":[{"id":"T9130","span":{"begin":925,"end":955},"obj":"Protein"},{"id":"T9129","span":{"begin":869,"end":917},"obj":"Protein"},{"id":"T9128","span":{"begin":304,"end":317},"obj":"Protein"},{"id":"T9127","span":{"begin":793,"end":796},"obj":"Entity"},{"id":"T9126","span":{"begin":975,"end":1004},"obj":"Protein"},{"id":"T9125","span":{"begin":1187,"end":1193},"obj":"Protein"},{"id":"T9124","span":{"begin":837,"end":844},"obj":"Protein"},{"id":"T9123","span":{"begin":853,"end":864},"obj":"Protein"},{"id":"T9122","span":{"begin":958,"end":973},"obj":"Protein"},{"id":"T9121","span":{"begin":837,"end":840},"obj":"Protein"},{"id":"T9120","span":{"begin":761,"end":766},"obj":"Protein"},{"id":"T9119","span":{"begin":845,"end":852},"obj":"Entity"},{"id":"T9118","span":{"begin":498,"end":516},"obj":"Protein"},{"id":"T9117","span":{"begin":482,"end":486},"obj":"Entity"},{"id":"T9116","span":{"begin":740,"end":749},"obj":"Entity"},{"id":"T9115","span":{"begin":1455,"end":1475},"obj":"Protein"},{"id":"T9114","span":{"begin":600,"end":603},"obj":"Protein"},{"id":"T9113","span":{"begin":637,"end":660},"obj":"Entity"},{"id":"T9112","span":{"begin":1298,"end":1303},"obj":"Protein"},{"id":"T9111","span":{"begin":932,"end":935},"obj":"Protein"},{"id":"T9110","span":{"begin":1414,"end":1418},"obj":"Protein"},{"id":"T9109","span":{"begin":927,"end":931},"obj":"Protein"},{"id":"T9166","span":{"begin":1414,"end":1429},"obj":"Gene_expression"},{"id":"T9165","span":{"begin":1414,"end":1429},"obj":"Transcription"},{"id":"T9164","span":{"begin":149,"end":167},"obj":"Negative_regulation"},{"id":"T9163","span":{"begin":1481,"end":1494},"obj":"Entity"},{"id":"T9162","span":{"begin":781,"end":784},"obj":"Protein"},{"id":"T9161","span":{"begin":789,"end":808},"obj":"Entity"},{"id":"T9160","span":{"begin":1288,"end":1293},"obj":"Protein"},{"id":"T9159","span":{"begin":975,"end":1004},"obj":"Entity"},{"id":"T9158","span":{"begin":0,"end":4},"obj":"Entity"},{"id":"T9157","span":{"begin":1386,"end":1399},"obj":"Entity"},{"id":"T9156","span":{"begin":975,"end":1004},"obj":"Protein"},{"id":"T9155","span":{"begin":102,"end":111},"obj":"Entity"},{"id":"T9154","span":{"begin":1177,"end":1182},"obj":"Protein"},{"id":"T9153","span":{"begin":493,"end":516},"obj":"Entity"},{"id":"T9152","span":{"begin":204,"end":221},"obj":"Protein"},{"id":"T9151","span":{"begin":87,"end":117},"obj":"Entity"},{"id":"T9150","span":{"begin":940,"end":943},"obj":"Protein"},{"id":"T9149","span":{"begin":637,"end":660},"obj":"Entity"},{"id":"T9148","span":{"begin":911,"end":916},"obj":"Protein"},{"id":"T9147","span":{"begin":1481,"end":1486},"obj":"Protein"},{"id":"T9146","span":{"begin":774,"end":788},"obj":"Entity"},{"id":"T9145","span":{"begin":149,"end":153},"obj":"Protein"},{"id":"T9144","span":{"begin":619,"end":636},"obj":"Entity"},{"id":"T9143","span":{"begin":626,"end":635},"obj":"Entity"},{"id":"T9142","span":{"begin":809,"end":818},"obj":"Protein"},{"id":"T9141","span":{"begin":299,"end":317},"obj":"Protein"},{"id":"T9140","span":{"begin":498,"end":501},"obj":"Protein"},{"id":"T9139","span":{"begin":1458,"end":1463},"obj":"Protein"},{"id":"T9138","span":{"begin":416,"end":419},"obj":"Protein"},{"id":"T9137","span":{"begin":1235,"end":1238},"obj":"Protein"},{"id":"T9136","span":{"begin":1079,"end":1096},"obj":"Protein"},{"id":"T9135","span":{"begin":48,"end":59},"obj":"Protein"},{"id":"T9134","span":{"begin":1039,"end":1042},"obj":"Entity"},{"id":"T9133","span":{"begin":789,"end":808},"obj":"Protein"}],"relations":[{"id":"R4819","pred":"themeOf","subj":"T9110","obj":"T9165"},{"id":"R4820","pred":"themeOf","subj":"T9110","obj":"T9166"},{"id":"R4821","pred":"themeOf","subj":"T9145","obj":"T9164"}],"text":"PBMC were incubated with various concentrations of anti-CD3 in the presence or absence of inhibitors (LY294002, PDTC). After 16 hours of incubation, mRNA was extracted with RNAzol B (Biotex Laboratories, Houston, TX, USA) in accordance with the manufacturer's instructions. Reverse transcription of 2 μg of total mRNA was performed at 42°C using the Superscript™ reverse transcription system (Takara, Shiga, Japan). PCR amplification of cDNA aliquots was performed by adding 2.5 mM dNTPs, 2.5 U of Taq DNA polymerase (Takara) and 0.25 μM of sense and antisense primers. The reaction was performed in PCR buffer (1.5 mM MgCl2, 50 mM KCl, 10 mM Tris-HCl, pH 8.3) in a total volume of 25 μl. The following sense and antisense primers for each molecules were used: IL-17 sense, 5'-ATG ACT CCT GGG AAG ACC TCA TTG-3'; IL-17 antisense, 5'-TTA GGC CAC ATG GTG GAC AAT CGG-3'; glyceraldehyde-3-phosphate dehydrogenase (GAPDH) sense, 5'-CGA TGC TGG GCG TGA GTA C-3'; GAPDH antisense, 5'-CGT TCA GCT CAG GGA TGA CC-3'. Reactions were processed in a DNA thermal cycler (Perkin-Elmer Cetus, Norwalk, CT, USA) through cycles for 30 s of denaturation at 94°C, 1 min of annealing at 56°C for GAPDH and IL-17, followed by 1 min of elongation at 72°C. PCR rounds were repeated for 25 cycles each for both GAPDH and IL-17; this was determined as falling within the exponential phase of amplification for each molecule. The level of mRNA expression was presented as a ratio of IL-17 PCR product over GAPDH product."}

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were incubated with various concentrations of anti-CD3 in the presence or absence of inhibitors (LY294002, PDTC). After 16 hours of incubation, mRNA was extracted with RNAzol B (Biotex Laboratories, Houston, TX, USA) in accordance with the manufacturer's instructions. Reverse transcription of 2 μg of total mRNA was performed at 42°C using the Superscript™ reverse transcription system (Takara, Shiga, Japan). PCR amplification of cDNA aliquots was performed by adding 2.5 mM dNTPs, 2.5 U of Taq DNA polymerase (Takara) and 0.25 μM of sense and antisense primers. The reaction was performed in PCR buffer (1.5 mM MgCl2, 50 mM KCl, 10 mM Tris-HCl, pH 8.3) in a total volume of 25 μl. The following sense and antisense primers for each molecules were used: IL-17 sense, 5'-ATG ACT CCT GGG AAG ACC TCA TTG-3'; IL-17 antisense, 5'-TTA GGC CAC ATG GTG GAC AAT CGG-3'; glyceraldehyde-3-phosphate dehydrogenase (GAPDH) sense, 5'-CGA TGC TGG GCG TGA GTA C-3'; GAPDH antisense, 5'-CGT TCA GCT CAG GGA TGA CC-3'. Reactions were processed in a DNA thermal cycler (Perkin-Elmer Cetus, Norwalk, CT, USA) through cycles for 30 s of denaturation at 94°C, 1 min of annealing at 56°C for GAPDH and IL-17, followed by 1 min of elongation at 72°C. PCR rounds were repeated for 25 cycles each for both GAPDH and IL-17; this was determined as falling within the exponential phase of amplification for each molecule. The level of mRNA expression was presented as a ratio of IL-17 PCR product over GAPDH product."}

{"project":"bionlp-st-ge-2016-test-tees","denotations":[{"id":"T8697","span":{"begin":1487,"end":1494},"obj":"Gene_expression"},{"id":"T8696","span":{"begin":1481,"end":1486},"obj":"Protein"},{"id":"T8695","span":{"begin":1298,"end":1303},"obj":"Protein"},{"id":"T8694","span":{"begin":1288,"end":1293},"obj":"Protein"},{"id":"T8693","span":{"begin":1187,"end":1192},"obj":"Protein"},{"id":"T8692","span":{"begin":1177,"end":1182},"obj":"Protein"},{"id":"T8691","span":{"begin":958,"end":963},"obj":"Protein"},{"id":"T8690","span":{"begin":911,"end":916},"obj":"Protein"},{"id":"T8689","span":{"begin":869,"end":909},"obj":"Protein"},{"id":"T8688","span":{"begin":498,"end":516},"obj":"Protein"},{"id":"T8687","span":{"begin":51,"end":59},"obj":"Protein"}],"relations":[{"id":"R4481","pred":"themeOf","subj":"T8696","obj":"T8697"}],"text":"PBMC were incubated with various concentrations of anti-CD3 in the presence or absence of inhibitors (LY294002, PDTC). After 16 hours of incubation, mRNA was extracted with RNAzol B (Biotex Laboratories, Houston, TX, USA) in accordance with the manufacturer's instructions. Reverse transcription of 2 μg of total mRNA was performed at 42°C using the Superscript™ reverse transcription system (Takara, Shiga, Japan). PCR amplification of cDNA aliquots was performed by adding 2.5 mM dNTPs, 2.5 U of Taq DNA polymerase (Takara) and 0.25 μM of sense and antisense primers. The reaction was performed in PCR buffer (1.5 mM MgCl2, 50 mM KCl, 10 mM Tris-HCl, pH 8.3) in a total volume of 25 μl. The following sense and antisense primers for each molecules were used: IL-17 sense, 5'-ATG ACT CCT GGG AAG ACC TCA TTG-3'; IL-17 antisense, 5'-TTA GGC CAC ATG GTG GAC AAT CGG-3'; glyceraldehyde-3-phosphate dehydrogenase (GAPDH) sense, 5'-CGA TGC TGG GCG TGA GTA C-3'; GAPDH antisense, 5'-CGT TCA GCT CAG GGA TGA CC-3'. Reactions were processed in a DNA thermal cycler (Perkin-Elmer Cetus, Norwalk, CT, USA) through cycles for 30 s of denaturation at 94°C, 1 min of annealing at 56°C for GAPDH and IL-17, followed by 1 min of elongation at 72°C. PCR rounds were repeated for 25 cycles each for both GAPDH and IL-17; this was determined as falling within the exponential phase of amplification for each molecule. The level of mRNA expression was presented as a ratio of IL-17 PCR product over GAPDH product."}

{"project":"testone","denotations":[{"id":"T8193","span":{"begin":1414,"end":1429},"obj":"Transcription"},{"id":"T8192","span":{"begin":1481,"end":1486},"obj":"Protein"},{"id":"T8191","span":{"begin":1458,"end":1463},"obj":"Protein"},{"id":"T8190","span":{"begin":1298,"end":1303},"obj":"Protein"},{"id":"T8189","span":{"begin":1288,"end":1293},"obj":"Protein"},{"id":"T8188","span":{"begin":1187,"end":1192},"obj":"Protein"},{"id":"T8187","span":{"begin":1177,"end":1182},"obj":"Protein"},{"id":"T8186","span":{"begin":958,"end":963},"obj":"Protein"},{"id":"T8185","span":{"begin":911,"end":916},"obj":"Protein"},{"id":"T8184","span":{"begin":869,"end":909},"obj":"Protein"},{"id":"T8183","span":{"begin":761,"end":766},"obj":"Protein"},{"id":"T8182","span":{"begin":498,"end":501},"obj":"Protein"}],"relations":[{"id":"R4470","pred":"equivalentTo","subj":"T8185","obj":"T8184"},{"id":"R4471","pred":"themeOf","subj":"T8191","obj":"T8193"},{"id":"R4472","pred":"themeOf","subj":"T8192","obj":"T8193"}],"text":"PBMC were incubated with various concentrations of anti-CD3 in the presence or absence of inhibitors (LY294002, PDTC). After 16 hours of incubation, mRNA was extracted with RNAzol B (Biotex Laboratories, Houston, TX, USA) in accordance with the manufacturer's instructions. Reverse transcription of 2 μg of total mRNA was performed at 42°C using the Superscript™ reverse transcription system (Takara, Shiga, Japan). PCR amplification of cDNA aliquots was performed by adding 2.5 mM dNTPs, 2.5 U of Taq DNA polymerase (Takara) and 0.25 μM of sense and antisense primers. The reaction was performed in PCR buffer (1.5 mM MgCl2, 50 mM KCl, 10 mM Tris-HCl, pH 8.3) in a total volume of 25 μl. The following sense and antisense primers for each molecules were used: IL-17 sense, 5'-ATG ACT CCT GGG AAG ACC TCA TTG-3'; IL-17 antisense, 5'-TTA GGC CAC ATG GTG GAC AAT CGG-3'; glyceraldehyde-3-phosphate dehydrogenase (GAPDH) sense, 5'-CGA TGC TGG GCG TGA GTA C-3'; GAPDH antisense, 5'-CGT TCA GCT CAG GGA TGA CC-3'. Reactions were processed in a DNA thermal cycler (Perkin-Elmer Cetus, Norwalk, CT, USA) through cycles for 30 s of denaturation at 94°C, 1 min of annealing at 56°C for GAPDH and IL-17, followed by 1 min of elongation at 72°C. PCR rounds were repeated for 25 cycles each for both GAPDH and IL-17; this was determined as falling within the exponential phase of amplification for each molecule. The level of mRNA expression was presented as a ratio of IL-17 PCR product over GAPDH product."}