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test3

2_test

{"project":"2_test","denotations":[{"id":"22396652-11009421-98461852","span":{"begin":283,"end":285},"obj":"11009421"},{"id":"22396652-20530205-98461853","span":{"begin":400,"end":402},"obj":"20530205"},{"id":"22396652-21841782-98461854","span":{"begin":626,"end":628},"obj":"21841782"},{"id":"22396652-18591409-98461855","span":{"begin":860,"end":862},"obj":"18591409"},{"id":"22396652-15210742-98461856","span":{"begin":1829,"end":1831},"obj":"15210742"},{"id":"22396652-9566918-98461857","span":{"begin":1907,"end":1909},"obj":"9566918"}],"text":"To study the effect of endogenously expressed A20 on RIG-I-induced signaling in a more immunological relevant context, we performed further experiments in A20 deficient primary macrophages. Since A20 full knockout mice die prematurely as a result of severe multi-organ inflammation [37], we generated mice carrying a conditional A20 allele in which exon IV and exon V were flanked by two loxP sites [56]. Crossing these mice with transgenic mice expressing Cre recombinase under control of the lysozyme M promoter leads to specific deletion in myeloid cells and allowed us to generate myeloid cell specific A20 knockout mice [48]. To stimulate the RIG-I receptor, we transfected A20myel-KO BMDM and wild type control cells with minimal amounts of low molecular weight (LMW) poly(I:C), which is known to preferentially bind and activate RIG-I rather than MDA5 [57]. Of note, this concentration of poly(I:C) was unable to induce significant TLR3 dependent NF-κB and IRF3 activation or cytokine production (data not shown). As expected, poly(I:C) transfection induced the rapid expression of A20 in wild type, but not in A20myel-KO BMDM (figure 1B, upper panel). At early time points, slightly slower migrating forms of A20 were observed, indicating that A20 undergoes a yet unknown modification upon poly(I:C) transfection. Compared to wild type BMBM, A20 deficient cells showed enhanced NF-κB activation as indicated by increased phosphorylation and sustained degradation of IκBα (figure 1B). Furthermore, nuclear translocation of the p65 NF-κB subunit was enhanced in poly(I:C) transfected A20myel-KO BMDM, reaching a maximum at earlier time points compared to wild type cells (figure 1C). IRF3 is known to be activated upon phosphorylation of a series of carboxyl terminal serine residues by the IKK-like kinases TBK1 and IKKε [58], leading to its dimerization and subsequent translocation to the nucleus [59]. Using immunoblotting with an antibody directed against phosphorylated Ser396, maximum IRF3 phosphorylation was detected at earlier time points in A20myel-KO BMDM compared to wild type BMDM (figure 1B). Similar to p65, IRF3 nuclear translocation reached its maximum at an earlier time point in A20 deficient BMDM compared to wild type cells (figure 1C). NF-κB controls the expression of IL-6 and TNF, and NF-κB and IRF3 control the expression of IFNβ. In line with the enhanced activation of NF-κB and IRF3 as described above, A20myel-KO BMDM secreted increased amounts of IL-6, TNF and IFNβ (figure 1D). Similar results were obtained using peritoneal macrophages (data not shown). Together, these results demonstrate that A20 plays an important role in the negative regulation of RIG-I-induced NF-κB and IRF3 activation in primary macrophages."}

pmc-enju-pas

bionlp-st-ge-2016-coref

{"project":"bionlp-st-ge-2016-coref","denotations":[{"id":"T4891","span":{"begin":1690,"end":1694},"obj":"Antecedent"},{"id":"T4890","span":{"begin":1845,"end":1848},"obj":"Anaphor"}],"relations":[{"id":"R3329","pred":"boundBy","subj":"T4890","obj":"T4891"}],"namespaces":[{"prefix":"_base","uri":"https://bionlp.dbcls.jp/ontology/ge.owl#"}],"text":"To study the effect of endogenously expressed A20 on RIG-I-induced signaling in a more immunological relevant context, we performed further experiments in A20 deficient primary macrophages. Since A20 full knockout mice die prematurely as a result of severe multi-organ inflammation [37], we generated mice carrying a conditional A20 allele in which exon IV and exon V were flanked by two loxP sites [56]. Crossing these mice with transgenic mice expressing Cre recombinase under control of the lysozyme M promoter leads to specific deletion in myeloid cells and allowed us to generate myeloid cell specific A20 knockout mice [48]. To stimulate the RIG-I receptor, we transfected A20myel-KO BMDM and wild type control cells with minimal amounts of low molecular weight (LMW) poly(I:C), which is known to preferentially bind and activate RIG-I rather than MDA5 [57]. Of note, this concentration of poly(I:C) was unable to induce significant TLR3 dependent NF-κB and IRF3 activation or cytokine production (data not shown). As expected, poly(I:C) transfection induced the rapid expression of A20 in wild type, but not in A20myel-KO BMDM (figure 1B, upper panel). At early time points, slightly slower migrating forms of A20 were observed, indicating that A20 undergoes a yet unknown modification upon poly(I:C) transfection. Compared to wild type BMBM, A20 deficient cells showed enhanced NF-κB activation as indicated by increased phosphorylation and sustained degradation of IκBα (figure 1B). Furthermore, nuclear translocation of the p65 NF-κB subunit was enhanced in poly(I:C) transfected A20myel-KO BMDM, reaching a maximum at earlier time points compared to wild type cells (figure 1C). IRF3 is known to be activated upon phosphorylation of a series of carboxyl terminal serine residues by the IKK-like kinases TBK1 and IKKε [58], leading to its dimerization and subsequent translocation to the nucleus [59]. Using immunoblotting with an antibody directed against phosphorylated Ser396, maximum IRF3 phosphorylation was detected at earlier time points in A20myel-KO BMDM compared to wild type BMDM (figure 1B). Similar to p65, IRF3 nuclear translocation reached its maximum at an earlier time point in A20 deficient BMDM compared to wild type cells (figure 1C). NF-κB controls the expression of IL-6 and TNF, and NF-κB and IRF3 control the expression of IFNβ. In line with the enhanced activation of NF-κB and IRF3 as described above, A20myel-KO BMDM secreted increased amounts of IL-6, TNF and IFNβ (figure 1D). Similar results were obtained using peritoneal macrophages (data not shown). Together, these results demonstrate that A20 plays an important role in the negative regulation of RIG-I-induced NF-κB and IRF3 activation in primary macrophages."}

bionlp-st-ge-2016-test-proteins

{"project":"bionlp-st-ge-2016-test-proteins","denotations":[{"id":"T4944","span":{"begin":2716,"end":2720},"obj":"Protein"},{"id":"T4943","span":{"begin":2692,"end":2697},"obj":"Protein"},{"id":"T4942","span":{"begin":2498,"end":2502},"obj":"Protein"},{"id":"T4941","span":{"begin":2490,"end":2493},"obj":"Protein"},{"id":"T4940","span":{"begin":2484,"end":2488},"obj":"Protein"},{"id":"T4939","span":{"begin":2413,"end":2417},"obj":"Protein"},{"id":"T4938","span":{"begin":2357,"end":2361},"obj":"Protein"},{"id":"T4937","span":{"begin":2326,"end":2330},"obj":"Protein"},{"id":"T4936","span":{"begin":2307,"end":2310},"obj":"Protein"},{"id":"T4935","span":{"begin":2298,"end":2302},"obj":"Protein"},{"id":"T4934","span":{"begin":2130,"end":2134},"obj":"Protein"},{"id":"T4933","span":{"begin":2125,"end":2128},"obj":"Protein"},{"id":"T4932","span":{"begin":1998,"end":2002},"obj":"Protein"},{"id":"T4931","span":{"begin":1823,"end":1827},"obj":"Protein"},{"id":"T4930","span":{"begin":1814,"end":1818},"obj":"Protein"},{"id":"T4929","span":{"begin":1690,"end":1694},"obj":"Protein"},{"id":"T4928","span":{"begin":1534,"end":1537},"obj":"Protein"},{"id":"T4927","span":{"begin":1474,"end":1478},"obj":"Protein"},{"id":"T4926","span":{"begin":964,"end":968},"obj":"Protein"},{"id":"T4925","span":{"begin":939,"end":943},"obj":"Protein"},{"id":"T4924","span":{"begin":854,"end":858},"obj":"Protein"},{"id":"T4923","span":{"begin":836,"end":841},"obj":"Protein"},{"id":"T4922","span":{"begin":648,"end":653},"obj":"Protein"},{"id":"T4921","span":{"begin":53,"end":58},"obj":"Protein"}],"namespaces":[{"prefix":"_base","uri":"http://bionlp.dbcls.jp/ontology/ge.owl#"}],"text":"To study the effect of endogenously expressed A20 on RIG-I-induced signaling in a more immunological relevant context, we performed further experiments in A20 deficient primary macrophages. Since A20 full knockout mice die prematurely as a result of severe multi-organ inflammation [37], we generated mice carrying a conditional A20 allele in which exon IV and exon V were flanked by two loxP sites [56]. Crossing these mice with transgenic mice expressing Cre recombinase under control of the lysozyme M promoter leads to specific deletion in myeloid cells and allowed us to generate myeloid cell specific A20 knockout mice [48]. To stimulate the RIG-I receptor, we transfected A20myel-KO BMDM and wild type control cells with minimal amounts of low molecular weight (LMW) poly(I:C), which is known to preferentially bind and activate RIG-I rather than MDA5 [57]. Of note, this concentration of poly(I:C) was unable to induce significant TLR3 dependent NF-κB and IRF3 activation or cytokine production (data not shown). As expected, poly(I:C) transfection induced the rapid expression of A20 in wild type, but not in A20myel-KO BMDM (figure 1B, upper panel). At early time points, slightly slower migrating forms of A20 were observed, indicating that A20 undergoes a yet unknown modification upon poly(I:C) transfection. Compared to wild type BMBM, A20 deficient cells showed enhanced NF-κB activation as indicated by increased phosphorylation and sustained degradation of IκBα (figure 1B). Furthermore, nuclear translocation of the p65 NF-κB subunit was enhanced in poly(I:C) transfected A20myel-KO BMDM, reaching a maximum at earlier time points compared to wild type cells (figure 1C). IRF3 is known to be activated upon phosphorylation of a series of carboxyl terminal serine residues by the IKK-like kinases TBK1 and IKKε [58], leading to its dimerization and subsequent translocation to the nucleus [59]. Using immunoblotting with an antibody directed against phosphorylated Ser396, maximum IRF3 phosphorylation was detected at earlier time points in A20myel-KO BMDM compared to wild type BMDM (figure 1B). Similar to p65, IRF3 nuclear translocation reached its maximum at an earlier time point in A20 deficient BMDM compared to wild type cells (figure 1C). NF-κB controls the expression of IL-6 and TNF, and NF-κB and IRF3 control the expression of IFNβ. In line with the enhanced activation of NF-κB and IRF3 as described above, A20myel-KO BMDM secreted increased amounts of IL-6, TNF and IFNβ (figure 1D). Similar results were obtained using peritoneal macrophages (data not shown). Together, these results demonstrate that A20 plays an important role in the negative regulation of RIG-I-induced NF-κB and IRF3 activation in primary macrophages."}

bionlp-st-ge-2016-uniprot

UBERON-AE

{"project":"UBERON-AE","denotations":[{"id":"T4723","span":{"begin":263,"end":268},"obj":"http://purl.obolibrary.org/obo/UBERON_0000062"}],"text":"To study the effect of endogenously expressed A20 on RIG-I-induced signaling in a more immunological relevant context, we performed further experiments in A20 deficient primary macrophages. Since A20 full knockout mice die prematurely as a result of severe multi-organ inflammation [37], we generated mice carrying a conditional A20 allele in which exon IV and exon V were flanked by two loxP sites [56]. Crossing these mice with transgenic mice expressing Cre recombinase under control of the lysozyme M promoter leads to specific deletion in myeloid cells and allowed us to generate myeloid cell specific A20 knockout mice [48]. To stimulate the RIG-I receptor, we transfected A20myel-KO BMDM and wild type control cells with minimal amounts of low molecular weight (LMW) poly(I:C), which is known to preferentially bind and activate RIG-I rather than MDA5 [57]. Of note, this concentration of poly(I:C) was unable to induce significant TLR3 dependent NF-κB and IRF3 activation or cytokine production (data not shown). As expected, poly(I:C) transfection induced the rapid expression of A20 in wild type, but not in A20myel-KO BMDM (figure 1B, upper panel). At early time points, slightly slower migrating forms of A20 were observed, indicating that A20 undergoes a yet unknown modification upon poly(I:C) transfection. Compared to wild type BMBM, A20 deficient cells showed enhanced NF-κB activation as indicated by increased phosphorylation and sustained degradation of IκBα (figure 1B). Furthermore, nuclear translocation of the p65 NF-κB subunit was enhanced in poly(I:C) transfected A20myel-KO BMDM, reaching a maximum at earlier time points compared to wild type cells (figure 1C). IRF3 is known to be activated upon phosphorylation of a series of carboxyl terminal serine residues by the IKK-like kinases TBK1 and IKKε [58], leading to its dimerization and subsequent translocation to the nucleus [59]. Using immunoblotting with an antibody directed against phosphorylated Ser396, maximum IRF3 phosphorylation was detected at earlier time points in A20myel-KO BMDM compared to wild type BMDM (figure 1B). Similar to p65, IRF3 nuclear translocation reached its maximum at an earlier time point in A20 deficient BMDM compared to wild type cells (figure 1C). NF-κB controls the expression of IL-6 and TNF, and NF-κB and IRF3 control the expression of IFNβ. In line with the enhanced activation of NF-κB and IRF3 as described above, A20myel-KO BMDM secreted increased amounts of IL-6, TNF and IFNβ (figure 1D). Similar results were obtained using peritoneal macrophages (data not shown). Together, these results demonstrate that A20 plays an important role in the negative regulation of RIG-I-induced NF-κB and IRF3 activation in primary macrophages."}

GO-BP

{"project":"GO-BP","denotations":[{"id":"T4949","span":{"begin":2389,"end":2408},"obj":"http://purl.obolibrary.org/obo/GO_0051092"},{"id":"T4947","span":{"begin":67,"end":76},"obj":"http://purl.obolibrary.org/obo/GO_0023052"},{"id":"T4962","span":{"begin":2678,"end":2688},"obj":"http://purl.obolibrary.org/obo/GO_0065007"},{"id":"T4961","span":{"begin":1814,"end":1818},"obj":"http://purl.obolibrary.org/obo/GO_0008384"},{"id":"T4960","span":{"begin":1797,"end":1800},"obj":"http://purl.obolibrary.org/obo/GO_0008384"},{"id":"T4959","span":{"begin":1459,"end":1470},"obj":"http://purl.obolibrary.org/obo/GO_0009056"},{"id":"T4958","span":{"begin":2003,"end":2018},"obj":"http://purl.obolibrary.org/obo/GO_0016310"},{"id":"T4957","span":{"begin":1725,"end":1740},"obj":"http://purl.obolibrary.org/obo/GO_0016310"},{"id":"T4956","span":{"begin":1429,"end":1444},"obj":"http://purl.obolibrary.org/obo/GO_0016310"},{"id":"T4955","span":{"begin":983,"end":1002},"obj":"http://purl.obolibrary.org/obo/GO_0001816"},{"id":"T4954","span":{"begin":969,"end":1002},"obj":"http://purl.obolibrary.org/obo/GO_0001819"},{"id":"T4953","span":{"begin":969,"end":1002},"obj":"http://purl.obolibrary.org/obo/GO_1903557"},{"id":"T4952","span":{"begin":269,"end":281},"obj":"http://purl.obolibrary.org/obo/GO_0006954"}],"text":"To study the effect of endogenously expressed A20 on RIG-I-induced signaling in a more immunological relevant context, we performed further experiments in A20 deficient primary macrophages. Since A20 full knockout mice die prematurely as a result of severe multi-organ inflammation [37], we generated mice carrying a conditional A20 allele in which exon IV and exon V were flanked by two loxP sites [56]. Crossing these mice with transgenic mice expressing Cre recombinase under control of the lysozyme M promoter leads to specific deletion in myeloid cells and allowed us to generate myeloid cell specific A20 knockout mice [48]. To stimulate the RIG-I receptor, we transfected A20myel-KO BMDM and wild type control cells with minimal amounts of low molecular weight (LMW) poly(I:C), which is known to preferentially bind and activate RIG-I rather than MDA5 [57]. Of note, this concentration of poly(I:C) was unable to induce significant TLR3 dependent NF-κB and IRF3 activation or cytokine production (data not shown). As expected, poly(I:C) transfection induced the rapid expression of A20 in wild type, but not in A20myel-KO BMDM (figure 1B, upper panel). At early time points, slightly slower migrating forms of A20 were observed, indicating that A20 undergoes a yet unknown modification upon poly(I:C) transfection. Compared to wild type BMBM, A20 deficient cells showed enhanced NF-κB activation as indicated by increased phosphorylation and sustained degradation of IκBα (figure 1B). Furthermore, nuclear translocation of the p65 NF-κB subunit was enhanced in poly(I:C) transfected A20myel-KO BMDM, reaching a maximum at earlier time points compared to wild type cells (figure 1C). IRF3 is known to be activated upon phosphorylation of a series of carboxyl terminal serine residues by the IKK-like kinases TBK1 and IKKε [58], leading to its dimerization and subsequent translocation to the nucleus [59]. Using immunoblotting with an antibody directed against phosphorylated Ser396, maximum IRF3 phosphorylation was detected at earlier time points in A20myel-KO BMDM compared to wild type BMDM (figure 1B). Similar to p65, IRF3 nuclear translocation reached its maximum at an earlier time point in A20 deficient BMDM compared to wild type cells (figure 1C). NF-κB controls the expression of IL-6 and TNF, and NF-κB and IRF3 control the expression of IFNβ. In line with the enhanced activation of NF-κB and IRF3 as described above, A20myel-KO BMDM secreted increased amounts of IL-6, TNF and IFNβ (figure 1D). Similar results were obtained using peritoneal macrophages (data not shown). Together, these results demonstrate that A20 plays an important role in the negative regulation of RIG-I-induced NF-κB and IRF3 activation in primary macrophages."}

GO-MF

{"project":"GO-MF","denotations":[{"id":"T4967","span":{"begin":2484,"end":2488},"obj":"http://purl.obolibrary.org/obo/GO_0005138"},{"id":"T4966","span":{"begin":2298,"end":2302},"obj":"http://purl.obolibrary.org/obo/GO_0005138"},{"id":"T4965","span":{"begin":1941,"end":1949},"obj":"http://purl.obolibrary.org/obo/GO_0003823"},{"id":"T4964","span":{"begin":1814,"end":1818},"obj":"http://purl.obolibrary.org/obo/GO_0008384"},{"id":"T4963","span":{"begin":1797,"end":1800},"obj":"http://purl.obolibrary.org/obo/GO_0008384"}],"text":"To study the effect of endogenously expressed A20 on RIG-I-induced signaling in a more immunological relevant context, we performed further experiments in A20 deficient primary macrophages. Since A20 full knockout mice die prematurely as a result of severe multi-organ inflammation [37], we generated mice carrying a conditional A20 allele in which exon IV and exon V were flanked by two loxP sites [56]. Crossing these mice with transgenic mice expressing Cre recombinase under control of the lysozyme M promoter leads to specific deletion in myeloid cells and allowed us to generate myeloid cell specific A20 knockout mice [48]. To stimulate the RIG-I receptor, we transfected A20myel-KO BMDM and wild type control cells with minimal amounts of low molecular weight (LMW) poly(I:C), which is known to preferentially bind and activate RIG-I rather than MDA5 [57]. Of note, this concentration of poly(I:C) was unable to induce significant TLR3 dependent NF-κB and IRF3 activation or cytokine production (data not shown). As expected, poly(I:C) transfection induced the rapid expression of A20 in wild type, but not in A20myel-KO BMDM (figure 1B, upper panel). At early time points, slightly slower migrating forms of A20 were observed, indicating that A20 undergoes a yet unknown modification upon poly(I:C) transfection. Compared to wild type BMBM, A20 deficient cells showed enhanced NF-κB activation as indicated by increased phosphorylation and sustained degradation of IκBα (figure 1B). Furthermore, nuclear translocation of the p65 NF-κB subunit was enhanced in poly(I:C) transfected A20myel-KO BMDM, reaching a maximum at earlier time points compared to wild type cells (figure 1C). IRF3 is known to be activated upon phosphorylation of a series of carboxyl terminal serine residues by the IKK-like kinases TBK1 and IKKε [58], leading to its dimerization and subsequent translocation to the nucleus [59]. Using immunoblotting with an antibody directed against phosphorylated Ser396, maximum IRF3 phosphorylation was detected at earlier time points in A20myel-KO BMDM compared to wild type BMDM (figure 1B). Similar to p65, IRF3 nuclear translocation reached its maximum at an earlier time point in A20 deficient BMDM compared to wild type cells (figure 1C). NF-κB controls the expression of IL-6 and TNF, and NF-κB and IRF3 control the expression of IFNβ. In line with the enhanced activation of NF-κB and IRF3 as described above, A20myel-KO BMDM secreted increased amounts of IL-6, TNF and IFNβ (figure 1D). Similar results were obtained using peritoneal macrophages (data not shown). Together, these results demonstrate that A20 plays an important role in the negative regulation of RIG-I-induced NF-κB and IRF3 activation in primary macrophages."}

GO-CC

{"project":"GO-CC","denotations":[{"id":"T4976","span":{"begin":1941,"end":1949},"obj":"http://purl.obolibrary.org/obo/GO_0042571"},{"id":"T4975","span":{"begin":1941,"end":1949},"obj":"http://purl.obolibrary.org/obo/GO_0019815"},{"id":"T4974","span":{"begin":1898,"end":1905},"obj":"http://purl.obolibrary.org/obo/GO_0005634"},{"id":"T4973","span":{"begin":2246,"end":2251},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T4972","span":{"begin":1671,"end":1676},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T4971","span":{"begin":1364,"end":1369},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T4970","span":{"begin":717,"end":722},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T4969","span":{"begin":552,"end":557},"obj":"http://purl.obolibrary.org/obo/GO_0005623"}],"text":"To study the effect of endogenously expressed A20 on RIG-I-induced signaling in a more immunological relevant context, we performed further experiments in A20 deficient primary macrophages. Since A20 full knockout mice die prematurely as a result of severe multi-organ inflammation [37], we generated mice carrying a conditional A20 allele in which exon IV and exon V were flanked by two loxP sites [56]. Crossing these mice with transgenic mice expressing Cre recombinase under control of the lysozyme M promoter leads to specific deletion in myeloid cells and allowed us to generate myeloid cell specific A20 knockout mice [48]. To stimulate the RIG-I receptor, we transfected A20myel-KO BMDM and wild type control cells with minimal amounts of low molecular weight (LMW) poly(I:C), which is known to preferentially bind and activate RIG-I rather than MDA5 [57]. Of note, this concentration of poly(I:C) was unable to induce significant TLR3 dependent NF-κB and IRF3 activation or cytokine production (data not shown). As expected, poly(I:C) transfection induced the rapid expression of A20 in wild type, but not in A20myel-KO BMDM (figure 1B, upper panel). At early time points, slightly slower migrating forms of A20 were observed, indicating that A20 undergoes a yet unknown modification upon poly(I:C) transfection. Compared to wild type BMBM, A20 deficient cells showed enhanced NF-κB activation as indicated by increased phosphorylation and sustained degradation of IκBα (figure 1B). Furthermore, nuclear translocation of the p65 NF-κB subunit was enhanced in poly(I:C) transfected A20myel-KO BMDM, reaching a maximum at earlier time points compared to wild type cells (figure 1C). IRF3 is known to be activated upon phosphorylation of a series of carboxyl terminal serine residues by the IKK-like kinases TBK1 and IKKε [58], leading to its dimerization and subsequent translocation to the nucleus [59]. Using immunoblotting with an antibody directed against phosphorylated Ser396, maximum IRF3 phosphorylation was detected at earlier time points in A20myel-KO BMDM compared to wild type BMDM (figure 1B). Similar to p65, IRF3 nuclear translocation reached its maximum at an earlier time point in A20 deficient BMDM compared to wild type cells (figure 1C). NF-κB controls the expression of IL-6 and TNF, and NF-κB and IRF3 control the expression of IFNβ. In line with the enhanced activation of NF-κB and IRF3 as described above, A20myel-KO BMDM secreted increased amounts of IL-6, TNF and IFNβ (figure 1D). Similar results were obtained using peritoneal macrophages (data not shown). Together, these results demonstrate that A20 plays an important role in the negative regulation of RIG-I-induced NF-κB and IRF3 activation in primary macrophages."}

sentences

simple1

{"project":"simple1","denotations":[{"id":"T5082","span":{"begin":2716,"end":2720},"obj":"Protein"},{"id":"T5081","span":{"begin":2692,"end":2697},"obj":"Protein"},{"id":"T5080","span":{"begin":2498,"end":2502},"obj":"Protein"},{"id":"T5079","span":{"begin":2490,"end":2493},"obj":"Protein"},{"id":"T5078","span":{"begin":2484,"end":2488},"obj":"Protein"},{"id":"T5077","span":{"begin":2413,"end":2417},"obj":"Protein"},{"id":"T5076","span":{"begin":2357,"end":2361},"obj":"Protein"},{"id":"T5075","span":{"begin":2326,"end":2330},"obj":"Protein"},{"id":"T5074","span":{"begin":2307,"end":2310},"obj":"Protein"},{"id":"T5073","span":{"begin":2298,"end":2302},"obj":"Protein"},{"id":"T5072","span":{"begin":2130,"end":2134},"obj":"Protein"},{"id":"T5071","span":{"begin":2125,"end":2128},"obj":"Protein"},{"id":"T5070","span":{"begin":1998,"end":2002},"obj":"Protein"},{"id":"T5069","span":{"begin":1823,"end":1827},"obj":"Protein"},{"id":"T5068","span":{"begin":1814,"end":1818},"obj":"Protein"},{"id":"T5067","span":{"begin":1690,"end":1694},"obj":"Protein"},{"id":"T5066","span":{"begin":1534,"end":1537},"obj":"Protein"},{"id":"T5065","span":{"begin":1474,"end":1478},"obj":"Protein"},{"id":"T5064","span":{"begin":964,"end":968},"obj":"Protein"},{"id":"T5063","span":{"begin":939,"end":943},"obj":"Protein"},{"id":"T5062","span":{"begin":854,"end":858},"obj":"Protein"},{"id":"T5061","span":{"begin":836,"end":841},"obj":"Protein"},{"id":"T5060","span":{"begin":648,"end":653},"obj":"Protein"},{"id":"T5059","span":{"begin":53,"end":58},"obj":"Protein"}],"text":"To study the effect of endogenously expressed A20 on RIG-I-induced signaling in a more immunological relevant context, we performed further experiments in A20 deficient primary macrophages. Since A20 full knockout mice die prematurely as a result of severe multi-organ inflammation [37], we generated mice carrying a conditional A20 allele in which exon IV and exon V were flanked by two loxP sites [56]. Crossing these mice with transgenic mice expressing Cre recombinase under control of the lysozyme M promoter leads to specific deletion in myeloid cells and allowed us to generate myeloid cell specific A20 knockout mice [48]. To stimulate the RIG-I receptor, we transfected A20myel-KO BMDM and wild type control cells with minimal amounts of low molecular weight (LMW) poly(I:C), which is known to preferentially bind and activate RIG-I rather than MDA5 [57]. Of note, this concentration of poly(I:C) was unable to induce significant TLR3 dependent NF-κB and IRF3 activation or cytokine production (data not shown). As expected, poly(I:C) transfection induced the rapid expression of A20 in wild type, but not in A20myel-KO BMDM (figure 1B, upper panel). At early time points, slightly slower migrating forms of A20 were observed, indicating that A20 undergoes a yet unknown modification upon poly(I:C) transfection. Compared to wild type BMBM, A20 deficient cells showed enhanced NF-κB activation as indicated by increased phosphorylation and sustained degradation of IκBα (figure 1B). Furthermore, nuclear translocation of the p65 NF-κB subunit was enhanced in poly(I:C) transfected A20myel-KO BMDM, reaching a maximum at earlier time points compared to wild type cells (figure 1C). IRF3 is known to be activated upon phosphorylation of a series of carboxyl terminal serine residues by the IKK-like kinases TBK1 and IKKε [58], leading to its dimerization and subsequent translocation to the nucleus [59]. Using immunoblotting with an antibody directed against phosphorylated Ser396, maximum IRF3 phosphorylation was detected at earlier time points in A20myel-KO BMDM compared to wild type BMDM (figure 1B). Similar to p65, IRF3 nuclear translocation reached its maximum at an earlier time point in A20 deficient BMDM compared to wild type cells (figure 1C). NF-κB controls the expression of IL-6 and TNF, and NF-κB and IRF3 control the expression of IFNβ. In line with the enhanced activation of NF-κB and IRF3 as described above, A20myel-KO BMDM secreted increased amounts of IL-6, TNF and IFNβ (figure 1D). Similar results were obtained using peritoneal macrophages (data not shown). Together, these results demonstrate that A20 plays an important role in the negative regulation of RIG-I-induced NF-κB and IRF3 activation in primary macrophages."}

BioNLP16_DUT

BioNLP16_Messiy

DLUT931

bionlp-st-ge-2016-test-ihmc

bionlp-st-ge-2016-spacy-parsed

bionlp-st-ge-2016-test-tees

testone

PMC:3291650 / 12452-15207 JSONTXT

Annnotations TAB JSON ListView MergeView

PMC:3291650 / 12452-15207 JSON TXT