PMC:7100515 / 39720-41227 JSONTXT

{"project":"LitCovid-PD-FMA-UBERON","denotations":[{"id":"T350","span":{"begin":15,"end":22},"obj":"Body_part"},{"id":"T351","span":{"begin":63,"end":75},"obj":"Body_part"},{"id":"T352","span":{"begin":93,"end":98},"obj":"Body_part"},{"id":"T353","span":{"begin":166,"end":171},"obj":"Body_part"},{"id":"T354","span":{"begin":213,"end":225},"obj":"Body_part"},{"id":"T355","span":{"begin":777,"end":781},"obj":"Body_part"},{"id":"T356","span":{"begin":994,"end":1002},"obj":"Body_part"},{"id":"T357","span":{"begin":1050,"end":1058},"obj":"Body_part"},{"id":"T358","span":{"begin":1084,"end":1092},"obj":"Body_part"},{"id":"T359","span":{"begin":1239,"end":1247},"obj":"Body_part"},{"id":"T360","span":{"begin":1326,"end":1334},"obj":"Body_part"},{"id":"T361","span":{"begin":1447,"end":1455},"obj":"Body_part"}],"attributes":[{"id":"A350","pred":"fma_id","subj":"T350","obj":"http://purl.org/sig/ont/fma/fma67257"},{"id":"A351","pred":"fma_id","subj":"T351","obj":"http://purl.org/sig/ont/fma/fma62925"},{"id":"A352","pred":"fma_id","subj":"T352","obj":"http://purl.org/sig/ont/fma/fma68646"},{"id":"A353","pred":"fma_id","subj":"T353","obj":"http://purl.org/sig/ont/fma/fma68646"},{"id":"A354","pred":"fma_id","subj":"T354","obj":"http://purl.org/sig/ont/fma/fma62925"},{"id":"A355","pred":"fma_id","subj":"T355","obj":"http://purl.org/sig/ont/fma/fma62100"},{"id":"A356","pred":"fma_id","subj":"T356","obj":"http://purl.org/sig/ont/fma/fma67257"},{"id":"A357","pred":"fma_id","subj":"T357","obj":"http://purl.org/sig/ont/fma/fma62871"},{"id":"A358","pred":"fma_id","subj":"T358","obj":"http://purl.org/sig/ont/fma/fma67257"},{"id":"A359","pred":"fma_id","subj":"T359","obj":"http://purl.org/sig/ont/fma/fma62871"},{"id":"A360","pred":"fma_id","subj":"T360","obj":"http://purl.org/sig/ont/fma/fma67257"},{"id":"A361","pred":"fma_id","subj":"T361","obj":"http://purl.org/sig/ont/fma/fma62871"}],"text":"Detection of S protein of SARS-CoV-2 by western blot\nThe spike glycoprotein of SARS-CoV-2 in cells and on pseudovirions were detected by using western blot. Briefly, cells transfected with plasmids encoding spike glycoprotein of SARS-CoV-2, SARS-CoV, MERS-CoV, and MHV were lysed at 40 h post transfection by RIPA buffer (20 mM Tris-HCl pH 7.5, 150 mM NaCl, 1 mM EDTA, 0.1% SDS, 1% NP40, 1×protease inhibitor cocktail). To pellet down pseudovirions, the viral supernatants were centrifuged at 25,000 rpm for 2 h in a Beckman SW41 rotor at 4 °C through a 20% sucrose cushion, and virus pellets were resuspended into 30 μl RIPA buffer. The samples were boiled for 10 min and separated in a 10% SDS-PAGE gel and transferred to nitrocellulose filter membranes. After blocked by 5% milk, the membranes were blotted with primary antibodies, followed by incubated with horseradish peroxidase (HRP) conjugated secondary antibodies (1:5000) and visualized with Chemiluminescent Reagent (Bio-Rad). MHV S proteins were detected using polyclonal goat anti-MHV S antibody AO4 (1:2000); SARS-CoV S proteins were blotted with either polyclonal anti-SARS S1 antibodies T62 (1:2000) (Sinobiological Inc, Beijing, China) or mouse monoclonal against SARS S1 antibody MM02 (1:1000) (Sinobiological Inc, Beijing, China), MERS-CoV and SARS-CoV-2 S proteins were detected using mouse monoclonal anti-MERS S (1:1000) (Sinobiological Inc, Beijing, China) and anti-FLAG M2 antibody (1:1000) (Sigma, St. Louis, MO, USA), respectively."}

{"project":"LitCovid-PD-UBERON","denotations":[{"id":"T23","span":{"begin":777,"end":781},"obj":"Body_part"}],"attributes":[{"id":"A23","pred":"uberon_id","subj":"T23","obj":"http://purl.obolibrary.org/obo/UBERON_0001913"}],"text":"Detection of S protein of SARS-CoV-2 by western blot\nThe spike glycoprotein of SARS-CoV-2 in cells and on pseudovirions were detected by using western blot. Briefly, cells transfected with plasmids encoding spike glycoprotein of SARS-CoV-2, SARS-CoV, MERS-CoV, and MHV were lysed at 40 h post transfection by RIPA buffer (20 mM Tris-HCl pH 7.5, 150 mM NaCl, 1 mM EDTA, 0.1% SDS, 1% NP40, 1×protease inhibitor cocktail). To pellet down pseudovirions, the viral supernatants were centrifuged at 25,000 rpm for 2 h in a Beckman SW41 rotor at 4 °C through a 20% sucrose cushion, and virus pellets were resuspended into 30 μl RIPA buffer. The samples were boiled for 10 min and separated in a 10% SDS-PAGE gel and transferred to nitrocellulose filter membranes. After blocked by 5% milk, the membranes were blotted with primary antibodies, followed by incubated with horseradish peroxidase (HRP) conjugated secondary antibodies (1:5000) and visualized with Chemiluminescent Reagent (Bio-Rad). MHV S proteins were detected using polyclonal goat anti-MHV S antibody AO4 (1:2000); SARS-CoV S proteins were blotted with either polyclonal anti-SARS S1 antibodies T62 (1:2000) (Sinobiological Inc, Beijing, China) or mouse monoclonal against SARS S1 antibody MM02 (1:1000) (Sinobiological Inc, Beijing, China), MERS-CoV and SARS-CoV-2 S proteins were detected using mouse monoclonal anti-MERS S (1:1000) (Sinobiological Inc, Beijing, China) and anti-FLAG M2 antibody (1:1000) (Sigma, St. Louis, MO, USA), respectively."}

{"project":"LitCovid-PD-MONDO","denotations":[{"id":"T350","span":{"begin":26,"end":34},"obj":"Disease"},{"id":"T351","span":{"begin":79,"end":87},"obj":"Disease"},{"id":"T352","span":{"begin":229,"end":237},"obj":"Disease"},{"id":"T353","span":{"begin":241,"end":249},"obj":"Disease"},{"id":"T354","span":{"begin":374,"end":380},"obj":"Disease"},{"id":"T355","span":{"begin":1073,"end":1081},"obj":"Disease"},{"id":"T356","span":{"begin":1134,"end":1138},"obj":"Disease"},{"id":"T357","span":{"begin":1231,"end":1235},"obj":"Disease"},{"id":"T358","span":{"begin":1313,"end":1321},"obj":"Disease"}],"attributes":[{"id":"A350","pred":"mondo_id","subj":"T350","obj":"http://purl.obolibrary.org/obo/MONDO_0005091"},{"id":"A351","pred":"mondo_id","subj":"T351","obj":"http://purl.obolibrary.org/obo/MONDO_0005091"},{"id":"A352","pred":"mondo_id","subj":"T352","obj":"http://purl.obolibrary.org/obo/MONDO_0005091"},{"id":"A353","pred":"mondo_id","subj":"T353","obj":"http://purl.obolibrary.org/obo/MONDO_0005091"},{"id":"A354","pred":"mondo_id","subj":"T354","obj":"http://purl.obolibrary.org/obo/MONDO_0044204"},{"id":"A355","pred":"mondo_id","subj":"T355","obj":"http://purl.obolibrary.org/obo/MONDO_0005091"},{"id":"A356","pred":"mondo_id","subj":"T356","obj":"http://purl.obolibrary.org/obo/MONDO_0005091"},{"id":"A357","pred":"mondo_id","subj":"T357","obj":"http://purl.obolibrary.org/obo/MONDO_0005091"},{"id":"A358","pred":"mondo_id","subj":"T358","obj":"http://purl.obolibrary.org/obo/MONDO_0005091"}],"text":"Detection of S protein of SARS-CoV-2 by western blot\nThe spike glycoprotein of SARS-CoV-2 in cells and on pseudovirions were detected by using western blot. Briefly, cells transfected with plasmids encoding spike glycoprotein of SARS-CoV-2, SARS-CoV, MERS-CoV, and MHV were lysed at 40 h post transfection by RIPA buffer (20 mM Tris-HCl pH 7.5, 150 mM NaCl, 1 mM EDTA, 0.1% SDS, 1% NP40, 1×protease inhibitor cocktail). To pellet down pseudovirions, the viral supernatants were centrifuged at 25,000 rpm for 2 h in a Beckman SW41 rotor at 4 °C through a 20% sucrose cushion, and virus pellets were resuspended into 30 μl RIPA buffer. The samples were boiled for 10 min and separated in a 10% SDS-PAGE gel and transferred to nitrocellulose filter membranes. After blocked by 5% milk, the membranes were blotted with primary antibodies, followed by incubated with horseradish peroxidase (HRP) conjugated secondary antibodies (1:5000) and visualized with Chemiluminescent Reagent (Bio-Rad). MHV S proteins were detected using polyclonal goat anti-MHV S antibody AO4 (1:2000); SARS-CoV S proteins were blotted with either polyclonal anti-SARS S1 antibodies T62 (1:2000) (Sinobiological Inc, Beijing, China) or mouse monoclonal against SARS S1 antibody MM02 (1:1000) (Sinobiological Inc, Beijing, China), MERS-CoV and SARS-CoV-2 S proteins were detected using mouse monoclonal anti-MERS S (1:1000) (Sinobiological Inc, Beijing, China) and anti-FLAG M2 antibody (1:1000) (Sigma, St. Louis, MO, USA), respectively."}

{"project":"LitCovid-PD-CLO","denotations":[{"id":"T889","span":{"begin":93,"end":98},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T890","span":{"begin":166,"end":171},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T891","span":{"begin":515,"end":516},"obj":"http://purl.obolibrary.org/obo/CLO_0001020"},{"id":"T892","span":{"begin":539,"end":543},"obj":"http://purl.obolibrary.org/obo/CLO_0001387"},{"id":"T893","span":{"begin":552,"end":556},"obj":"http://purl.obolibrary.org/obo/CLO_0001564"},{"id":"T894","span":{"begin":552,"end":556},"obj":"http://purl.obolibrary.org/obo/CLO_0050194"},{"id":"T895","span":{"begin":579,"end":584},"obj":"http://purl.obolibrary.org/obo/NCBITaxon_10239"},{"id":"T896","span":{"begin":686,"end":690},"obj":"http://purl.obolibrary.org/obo/CLO_0001550"},{"id":"T897","span":{"begin":746,"end":755},"obj":"http://purl.obolibrary.org/obo/UBERON_0000158"},{"id":"T898","span":{"begin":787,"end":796},"obj":"http://purl.obolibrary.org/obo/UBERON_0000158"},{"id":"T899","span":{"begin":1034,"end":1038},"obj":"http://purl.obolibrary.org/obo/NCBITaxon_9925"},{"id":"T900","span":{"begin":1139,"end":1141},"obj":"http://purl.obolibrary.org/obo/CLO_0050050"},{"id":"T901","span":{"begin":1206,"end":1211},"obj":"http://purl.obolibrary.org/obo/CLO_0007836"},{"id":"T902","span":{"begin":1236,"end":1238},"obj":"http://purl.obolibrary.org/obo/CLO_0050050"},{"id":"T903","span":{"begin":1355,"end":1360},"obj":"http://purl.obolibrary.org/obo/CLO_0007836"},{"id":"T904","span":{"begin":1473,"end":1475},"obj":"http://purl.obolibrary.org/obo/CLO_0009141"},{"id":"T905","span":{"begin":1473,"end":1475},"obj":"http://purl.obolibrary.org/obo/CLO_0050980"},{"id":"T906","span":{"begin":1484,"end":1486},"obj":"http://purl.obolibrary.org/obo/CLO_0007815"}],"text":"Detection of S protein of SARS-CoV-2 by western blot\nThe spike glycoprotein of SARS-CoV-2 in cells and on pseudovirions were detected by using western blot. Briefly, cells transfected with plasmids encoding spike glycoprotein of SARS-CoV-2, SARS-CoV, MERS-CoV, and MHV were lysed at 40 h post transfection by RIPA buffer (20 mM Tris-HCl pH 7.5, 150 mM NaCl, 1 mM EDTA, 0.1% SDS, 1% NP40, 1×protease inhibitor cocktail). To pellet down pseudovirions, the viral supernatants were centrifuged at 25,000 rpm for 2 h in a Beckman SW41 rotor at 4 °C through a 20% sucrose cushion, and virus pellets were resuspended into 30 μl RIPA buffer. The samples were boiled for 10 min and separated in a 10% SDS-PAGE gel and transferred to nitrocellulose filter membranes. After blocked by 5% milk, the membranes were blotted with primary antibodies, followed by incubated with horseradish peroxidase (HRP) conjugated secondary antibodies (1:5000) and visualized with Chemiluminescent Reagent (Bio-Rad). MHV S proteins were detected using polyclonal goat anti-MHV S antibody AO4 (1:2000); SARS-CoV S proteins were blotted with either polyclonal anti-SARS S1 antibodies T62 (1:2000) (Sinobiological Inc, Beijing, China) or mouse monoclonal against SARS S1 antibody MM02 (1:1000) (Sinobiological Inc, Beijing, China), MERS-CoV and SARS-CoV-2 S proteins were detected using mouse monoclonal anti-MERS S (1:1000) (Sinobiological Inc, Beijing, China) and anti-FLAG M2 antibody (1:1000) (Sigma, St. Louis, MO, USA), respectively."}

{"project":"LitCovid-PD-CHEBI","denotations":[{"id":"T333","span":{"begin":15,"end":22},"obj":"Chemical"},{"id":"T334","span":{"begin":63,"end":75},"obj":"Chemical"},{"id":"T335","span":{"begin":213,"end":225},"obj":"Chemical"},{"id":"T336","span":{"begin":314,"end":320},"obj":"Chemical"},{"id":"T337","span":{"begin":328,"end":332},"obj":"Chemical"},{"id":"T338","span":{"begin":333,"end":336},"obj":"Chemical"},{"id":"T339","span":{"begin":352,"end":356},"obj":"Chemical"},{"id":"T340","span":{"begin":363,"end":367},"obj":"Chemical"},{"id":"T342","span":{"begin":374,"end":377},"obj":"Chemical"},{"id":"T343","span":{"begin":382,"end":386},"obj":"Chemical"},{"id":"T344","span":{"begin":390,"end":408},"obj":"Chemical"},{"id":"T346","span":{"begin":399,"end":408},"obj":"Chemical"},{"id":"T347","span":{"begin":558,"end":565},"obj":"Chemical"},{"id":"T348","span":{"begin":626,"end":632},"obj":"Chemical"},{"id":"T349","span":{"begin":692,"end":695},"obj":"Chemical"},{"id":"T350","span":{"begin":724,"end":738},"obj":"Chemical"},{"id":"T351","span":{"begin":994,"end":1002},"obj":"Chemical"},{"id":"T352","span":{"begin":1084,"end":1092},"obj":"Chemical"},{"id":"T353","span":{"begin":1326,"end":1334},"obj":"Chemical"},{"id":"T354","span":{"begin":1444,"end":1446},"obj":"Chemical"}],"attributes":[{"id":"A333","pred":"chebi_id","subj":"T333","obj":"http://purl.obolibrary.org/obo/CHEBI_36080"},{"id":"A334","pred":"chebi_id","subj":"T334","obj":"http://purl.obolibrary.org/obo/CHEBI_17089"},{"id":"A335","pred":"chebi_id","subj":"T335","obj":"http://purl.obolibrary.org/obo/CHEBI_17089"},{"id":"A336","pred":"chebi_id","subj":"T336","obj":"http://purl.obolibrary.org/obo/CHEBI_35225"},{"id":"A337","pred":"chebi_id","subj":"T337","obj":"http://purl.obolibrary.org/obo/CHEBI_9754"},{"id":"A338","pred":"chebi_id","subj":"T338","obj":"http://purl.obolibrary.org/obo/CHEBI_17883"},{"id":"A339","pred":"chebi_id","subj":"T339","obj":"http://purl.obolibrary.org/obo/CHEBI_26710"},{"id":"A340","pred":"chebi_id","subj":"T340","obj":"http://purl.obolibrary.org/obo/CHEBI_42191"},{"id":"A341","pred":"chebi_id","subj":"T340","obj":"http://purl.obolibrary.org/obo/CHEBI_64755"},{"id":"A342","pred":"chebi_id","subj":"T342","obj":"http://purl.obolibrary.org/obo/CHEBI_8984"},{"id":"A343","pred":"chebi_id","subj":"T343","obj":"http://purl.obolibrary.org/obo/CHEBI_63016"},{"id":"A344","pred":"chebi_id","subj":"T344","obj":"http://purl.obolibrary.org/obo/CHEBI_37670"},{"id":"A345","pred":"chebi_id","subj":"T344","obj":"http://purl.obolibrary.org/obo/CHEBI_60258"},{"id":"A346","pred":"chebi_id","subj":"T346","obj":"http://purl.obolibrary.org/obo/CHEBI_35222"},{"id":"A347","pred":"chebi_id","subj":"T347","obj":"http://purl.obolibrary.org/obo/CHEBI_17992"},{"id":"A348","pred":"chebi_id","subj":"T348","obj":"http://purl.obolibrary.org/obo/CHEBI_35225"},{"id":"A349","pred":"chebi_id","subj":"T349","obj":"http://purl.obolibrary.org/obo/CHEBI_8984"},{"id":"A350","pred":"chebi_id","subj":"T350","obj":"http://purl.obolibrary.org/obo/CHEBI_53325"},{"id":"A351","pred":"chebi_id","subj":"T351","obj":"http://purl.obolibrary.org/obo/CHEBI_36080"},{"id":"A352","pred":"chebi_id","subj":"T352","obj":"http://purl.obolibrary.org/obo/CHEBI_36080"},{"id":"A353","pred":"chebi_id","subj":"T353","obj":"http://purl.obolibrary.org/obo/CHEBI_36080"},{"id":"A354","pred":"chebi_id","subj":"T354","obj":"http://purl.obolibrary.org/obo/CHEBI_34827"},{"id":"A355","pred":"chebi_id","subj":"T354","obj":"http://purl.obolibrary.org/obo/CHEBI_51112"}],"text":"Detection of S protein of SARS-CoV-2 by western blot\nThe spike glycoprotein of SARS-CoV-2 in cells and on pseudovirions were detected by using western blot. Briefly, cells transfected with plasmids encoding spike glycoprotein of SARS-CoV-2, SARS-CoV, MERS-CoV, and MHV were lysed at 40 h post transfection by RIPA buffer (20 mM Tris-HCl pH 7.5, 150 mM NaCl, 1 mM EDTA, 0.1% SDS, 1% NP40, 1×protease inhibitor cocktail). To pellet down pseudovirions, the viral supernatants were centrifuged at 25,000 rpm for 2 h in a Beckman SW41 rotor at 4 °C through a 20% sucrose cushion, and virus pellets were resuspended into 30 μl RIPA buffer. The samples were boiled for 10 min and separated in a 10% SDS-PAGE gel and transferred to nitrocellulose filter membranes. After blocked by 5% milk, the membranes were blotted with primary antibodies, followed by incubated with horseradish peroxidase (HRP) conjugated secondary antibodies (1:5000) and visualized with Chemiluminescent Reagent (Bio-Rad). MHV S proteins were detected using polyclonal goat anti-MHV S antibody AO4 (1:2000); SARS-CoV S proteins were blotted with either polyclonal anti-SARS S1 antibodies T62 (1:2000) (Sinobiological Inc, Beijing, China) or mouse monoclonal against SARS S1 antibody MM02 (1:1000) (Sinobiological Inc, Beijing, China), MERS-CoV and SARS-CoV-2 S proteins were detected using mouse monoclonal anti-MERS S (1:1000) (Sinobiological Inc, Beijing, China) and anti-FLAG M2 antibody (1:1000) (Sigma, St. Louis, MO, USA), respectively."}

{"project":"LitCovid-PD-GO-BP","denotations":[{"id":"T79","span":{"begin":862,"end":890},"obj":"http://purl.obolibrary.org/obo/GO_0004601"}],"text":"Detection of S protein of SARS-CoV-2 by western blot\nThe spike glycoprotein of SARS-CoV-2 in cells and on pseudovirions were detected by using western blot. Briefly, cells transfected with plasmids encoding spike glycoprotein of SARS-CoV-2, SARS-CoV, MERS-CoV, and MHV were lysed at 40 h post transfection by RIPA buffer (20 mM Tris-HCl pH 7.5, 150 mM NaCl, 1 mM EDTA, 0.1% SDS, 1% NP40, 1×protease inhibitor cocktail). To pellet down pseudovirions, the viral supernatants were centrifuged at 25,000 rpm for 2 h in a Beckman SW41 rotor at 4 °C through a 20% sucrose cushion, and virus pellets were resuspended into 30 μl RIPA buffer. The samples were boiled for 10 min and separated in a 10% SDS-PAGE gel and transferred to nitrocellulose filter membranes. After blocked by 5% milk, the membranes were blotted with primary antibodies, followed by incubated with horseradish peroxidase (HRP) conjugated secondary antibodies (1:5000) and visualized with Chemiluminescent Reagent (Bio-Rad). MHV S proteins were detected using polyclonal goat anti-MHV S antibody AO4 (1:2000); SARS-CoV S proteins were blotted with either polyclonal anti-SARS S1 antibodies T62 (1:2000) (Sinobiological Inc, Beijing, China) or mouse monoclonal against SARS S1 antibody MM02 (1:1000) (Sinobiological Inc, Beijing, China), MERS-CoV and SARS-CoV-2 S proteins were detected using mouse monoclonal anti-MERS S (1:1000) (Sinobiological Inc, Beijing, China) and anti-FLAG M2 antibody (1:1000) (Sigma, St. Louis, MO, USA), respectively."}

{"project":"LitCovid-sentences","denotations":[{"id":"T286","span":{"begin":0,"end":52},"obj":"Sentence"},{"id":"T287","span":{"begin":53,"end":156},"obj":"Sentence"},{"id":"T288","span":{"begin":157,"end":419},"obj":"Sentence"},{"id":"T289","span":{"begin":420,"end":633},"obj":"Sentence"},{"id":"T290","span":{"begin":634,"end":756},"obj":"Sentence"},{"id":"T291","span":{"begin":757,"end":987},"obj":"Sentence"},{"id":"T292","span":{"begin":988,"end":1507},"obj":"Sentence"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/ontology/tao.owl#"}],"text":"Detection of S protein of SARS-CoV-2 by western blot\nThe spike glycoprotein of SARS-CoV-2 in cells and on pseudovirions were detected by using western blot. Briefly, cells transfected with plasmids encoding spike glycoprotein of SARS-CoV-2, SARS-CoV, MERS-CoV, and MHV were lysed at 40 h post transfection by RIPA buffer (20 mM Tris-HCl pH 7.5, 150 mM NaCl, 1 mM EDTA, 0.1% SDS, 1% NP40, 1×protease inhibitor cocktail). To pellet down pseudovirions, the viral supernatants were centrifuged at 25,000 rpm for 2 h in a Beckman SW41 rotor at 4 °C through a 20% sucrose cushion, and virus pellets were resuspended into 30 μl RIPA buffer. The samples were boiled for 10 min and separated in a 10% SDS-PAGE gel and transferred to nitrocellulose filter membranes. After blocked by 5% milk, the membranes were blotted with primary antibodies, followed by incubated with horseradish peroxidase (HRP) conjugated secondary antibodies (1:5000) and visualized with Chemiluminescent Reagent (Bio-Rad). MHV S proteins were detected using polyclonal goat anti-MHV S antibody AO4 (1:2000); SARS-CoV S proteins were blotted with either polyclonal anti-SARS S1 antibodies T62 (1:2000) (Sinobiological Inc, Beijing, China) or mouse monoclonal against SARS S1 antibody MM02 (1:1000) (Sinobiological Inc, Beijing, China), MERS-CoV and SARS-CoV-2 S proteins were detected using mouse monoclonal anti-MERS S (1:1000) (Sinobiological Inc, Beijing, China) and anti-FLAG M2 antibody (1:1000) (Sigma, St. Louis, MO, USA), respectively."}

{"project":"LitCovid-PubTator","denotations":[{"id":"1604","span":{"begin":13,"end":14},"obj":"Gene"},{"id":"1605","span":{"begin":26,"end":36},"obj":"Species"},{"id":"1628","span":{"begin":1324,"end":1325},"obj":"Gene"},{"id":"1629","span":{"begin":79,"end":89},"obj":"Species"},{"id":"1630","span":{"begin":229,"end":239},"obj":"Species"},{"id":"1631","span":{"begin":241,"end":249},"obj":"Species"},{"id":"1632","span":{"begin":251,"end":259},"obj":"Species"},{"id":"1633","span":{"begin":862,"end":873},"obj":"Species"},{"id":"1634","span":{"begin":988,"end":993},"obj":"Species"},{"id":"1635","span":{"begin":1073,"end":1081},"obj":"Species"},{"id":"1636","span":{"begin":1300,"end":1308},"obj":"Species"},{"id":"1637","span":{"begin":1313,"end":1323},"obj":"Species"},{"id":"1638","span":{"begin":265,"end":268},"obj":"Species"},{"id":"1639","span":{"begin":1044,"end":1047},"obj":"Species"},{"id":"1640","span":{"begin":1206,"end":1211},"obj":"Species"},{"id":"1641","span":{"begin":1355,"end":1360},"obj":"Species"},{"id":"1642","span":{"begin":309,"end":320},"obj":"Chemical"},{"id":"1643","span":{"begin":328,"end":332},"obj":"Chemical"},{"id":"1644","span":{"begin":333,"end":336},"obj":"Chemical"},{"id":"1645","span":{"begin":352,"end":356},"obj":"Chemical"},{"id":"1646","span":{"begin":363,"end":367},"obj":"Chemical"},{"id":"1647","span":{"begin":374,"end":377},"obj":"Chemical"},{"id":"1648","span":{"begin":558,"end":565},"obj":"Chemical"},{"id":"1649","span":{"begin":692,"end":695},"obj":"Chemical"}],"attributes":[{"id":"A1604","pred":"tao:has_database_id","subj":"1604","obj":"Gene:43740568"},{"id":"A1605","pred":"tao:has_database_id","subj":"1605","obj":"Tax:2697049"},{"id":"A1628","pred":"tao:has_database_id","subj":"1628","obj":"Gene:43740568"},{"id":"A1629","pred":"tao:has_database_id","subj":"1629","obj":"Tax:2697049"},{"id":"A1630","pred":"tao:has_database_id","subj":"1630","obj":"Tax:2697049"},{"id":"A1631","pred":"tao:has_database_id","subj":"1631","obj":"Tax:694009"},{"id":"A1632","pred":"tao:has_database_id","subj":"1632","obj":"Tax:1335626"},{"id":"A1633","pred":"tao:has_database_id","subj":"1633","obj":"Tax:3704"},{"id":"A1634","pred":"tao:has_database_id","subj":"1634","obj":"Tax:11145"},{"id":"A1635","pred":"tao:has_database_id","subj":"1635","obj":"Tax:694009"},{"id":"A1636","pred":"tao:has_database_id","subj":"1636","obj":"Tax:1335626"},{"id":"A1637","pred":"tao:has_database_id","subj":"1637","obj":"Tax:2697049"},{"id":"A1638","pred":"tao:has_database_id","subj":"1638","obj":"Tax:11138"},{"id":"A1639","pred":"tao:has_database_id","subj":"1639","obj":"Tax:11138"},{"id":"A1640","pred":"tao:has_database_id","subj":"1640","obj":"Tax:10090"},{"id":"A1641","pred":"tao:has_database_id","subj":"1641","obj":"Tax:10090"},{"id":"A1644","pred":"tao:has_database_id","subj":"1644","obj":"MESH:D006851"},{"id":"A1645","pred":"tao:has_database_id","subj":"1645","obj":"MESH:D012965"},{"id":"A1646","pred":"tao:has_database_id","subj":"1646","obj":"MESH:D004492"},{"id":"A1647","pred":"tao:has_database_id","subj":"1647","obj":"MESH:D012967"},{"id":"A1648","pred":"tao:has_database_id","subj":"1648","obj":"MESH:D013395"},{"id":"A1649","pred":"tao:has_database_id","subj":"1649","obj":"MESH:D012967"}],"namespaces":[{"prefix":"Tax","uri":"https://www.ncbi.nlm.nih.gov/taxonomy/"},{"prefix":"MESH","uri":"https://id.nlm.nih.gov/mesh/"},{"prefix":"Gene","uri":"https://www.ncbi.nlm.nih.gov/gene/"},{"prefix":"CVCL","uri":"https://web.expasy.org/cellosaurus/CVCL_"}],"text":"Detection of S protein of SARS-CoV-2 by western blot\nThe spike glycoprotein of SARS-CoV-2 in cells and on pseudovirions were detected by using western blot. Briefly, cells transfected with plasmids encoding spike glycoprotein of SARS-CoV-2, SARS-CoV, MERS-CoV, and MHV were lysed at 40 h post transfection by RIPA buffer (20 mM Tris-HCl pH 7.5, 150 mM NaCl, 1 mM EDTA, 0.1% SDS, 1% NP40, 1×protease inhibitor cocktail). To pellet down pseudovirions, the viral supernatants were centrifuged at 25,000 rpm for 2 h in a Beckman SW41 rotor at 4 °C through a 20% sucrose cushion, and virus pellets were resuspended into 30 μl RIPA buffer. The samples were boiled for 10 min and separated in a 10% SDS-PAGE gel and transferred to nitrocellulose filter membranes. After blocked by 5% milk, the membranes were blotted with primary antibodies, followed by incubated with horseradish peroxidase (HRP) conjugated secondary antibodies (1:5000) and visualized with Chemiluminescent Reagent (Bio-Rad). MHV S proteins were detected using polyclonal goat anti-MHV S antibody AO4 (1:2000); SARS-CoV S proteins were blotted with either polyclonal anti-SARS S1 antibodies T62 (1:2000) (Sinobiological Inc, Beijing, China) or mouse monoclonal against SARS S1 antibody MM02 (1:1000) (Sinobiological Inc, Beijing, China), MERS-CoV and SARS-CoV-2 S proteins were detected using mouse monoclonal anti-MERS S (1:1000) (Sinobiological Inc, Beijing, China) and anti-FLAG M2 antibody (1:1000) (Sigma, St. Louis, MO, USA), respectively."}