PMC:3589482 / 12598-13790 JSON TXT

Annnotations TAB JSON ListView MergeView

bionlp-st-ge-2016-spacy-parsed

pmc-enju-pas

bionlp-st-ge-2016-test-proteins

{"project":"bionlp-st-ge-2016-test-proteins","denotations":[{"id":"T6650","span":{"begin":1135,"end":1142},"obj":"Protein"},{"id":"T6649","span":{"begin":325,"end":332},"obj":"Protein"},{"id":"T6648","span":{"begin":204,"end":209},"obj":"Protein"},{"id":"T6647","span":{"begin":142,"end":147},"obj":"Protein"}],"namespaces":[{"prefix":"_base","uri":"http://bionlp.dbcls.jp/ontology/ge.owl#"}],"text":"Lentiviral shRNAs, Packaging and Transduction\npLKO.1-puro lentiviral vectors carrying short hairpin RNAs (shRNA) specific for human and mouse cebpb were used in these studies. Because the human and mouse cebpb 3′ untranslated regions are not identical, these species-specific shRNAs cannot knockdown expression of endogenous C/EBP-β when used on cells of the other species; therefore we used the human CEBPB shRNA as a control in these experiments as in [38]. To produce lentiviral particles, HEK-293T cells were transfected with a mixture of plasmids: each shRNA expression plasmid (1 µg), pCMV-ΔR8.2dvpr packaging plasmid (0.75 µg), and pCMV-VSV-G envelope plasmid (0.25 µg) using Lipofectamine 2000 reagent (Invitrogen). The lentiviral supernatant was collected 48 hrs after transfection, cleared by centrifugation at 2,000 g for 10 mins and passed through a 0.45 µm filter. The target cells were treated with the lentiviral supernatant and 8 µg/ml Polybrene (American Bioanalytical) for 24 hrs. The lentiviral supernatant was replaced with fresh growth media and incubated further for 72 hrs to allow for effective gene knockdown. C/EBP-β knockdown was confirmed by western blot analysis."}

bionlp-st-ge-2016-uniprot

{"project":"bionlp-st-ge-2016-uniprot","denotations":[{"id":"T6874","span":{"begin":1135,"end":1142},"obj":"http://www.uniprot.org/uniprot/P28033"},{"id":"T6873","span":{"begin":325,"end":332},"obj":"http://www.uniprot.org/uniprot/P28033"},{"id":"T6872","span":{"begin":204,"end":209},"obj":"http://www.uniprot.org/uniprot/P28033"},{"id":"T6871","span":{"begin":142,"end":147},"obj":"http://www.uniprot.org/uniprot/P28033"}],"namespaces":[{"prefix":"_base","uri":"http://www.uniprot.org/uniprot/"}],"text":"Lentiviral shRNAs, Packaging and Transduction\npLKO.1-puro lentiviral vectors carrying short hairpin RNAs (shRNA) specific for human and mouse cebpb were used in these studies. Because the human and mouse cebpb 3′ untranslated regions are not identical, these species-specific shRNAs cannot knockdown expression of endogenous C/EBP-β when used on cells of the other species; therefore we used the human CEBPB shRNA as a control in these experiments as in [38]. To produce lentiviral particles, HEK-293T cells were transfected with a mixture of plasmids: each shRNA expression plasmid (1 µg), pCMV-ΔR8.2dvpr packaging plasmid (0.75 µg), and pCMV-VSV-G envelope plasmid (0.25 µg) using Lipofectamine 2000 reagent (Invitrogen). The lentiviral supernatant was collected 48 hrs after transfection, cleared by centrifugation at 2,000 g for 10 mins and passed through a 0.45 µm filter. The target cells were treated with the lentiviral supernatant and 8 µg/ml Polybrene (American Bioanalytical) for 24 hrs. The lentiviral supernatant was replaced with fresh growth media and incubated further for 72 hrs to allow for effective gene knockdown. C/EBP-β knockdown was confirmed by western blot analysis."}

GO-BP

{"project":"GO-BP","denotations":[{"id":"T6652","span":{"begin":1050,"end":1056},"obj":"http://purl.obolibrary.org/obo/GO_0040007"},{"id":"T6651","span":{"begin":33,"end":45},"obj":"http://purl.obolibrary.org/obo/GO_0009293"}],"text":"Lentiviral shRNAs, Packaging and Transduction\npLKO.1-puro lentiviral vectors carrying short hairpin RNAs (shRNA) specific for human and mouse cebpb were used in these studies. Because the human and mouse cebpb 3′ untranslated regions are not identical, these species-specific shRNAs cannot knockdown expression of endogenous C/EBP-β when used on cells of the other species; therefore we used the human CEBPB shRNA as a control in these experiments as in [38]. To produce lentiviral particles, HEK-293T cells were transfected with a mixture of plasmids: each shRNA expression plasmid (1 µg), pCMV-ΔR8.2dvpr packaging plasmid (0.75 µg), and pCMV-VSV-G envelope plasmid (0.25 µg) using Lipofectamine 2000 reagent (Invitrogen). The lentiviral supernatant was collected 48 hrs after transfection, cleared by centrifugation at 2,000 g for 10 mins and passed through a 0.45 µm filter. The target cells were treated with the lentiviral supernatant and 8 µg/ml Polybrene (American Bioanalytical) for 24 hrs. The lentiviral supernatant was replaced with fresh growth media and incubated further for 72 hrs to allow for effective gene knockdown. C/EBP-β knockdown was confirmed by western blot analysis."}

GO-CC

{"project":"GO-CC","denotations":[{"id":"T6656","span":{"begin":650,"end":658},"obj":"http://purl.obolibrary.org/obo/GO_0009274"},{"id":"T6655","span":{"begin":889,"end":894},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T6654","span":{"begin":502,"end":507},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T6653","span":{"begin":346,"end":351},"obj":"http://purl.obolibrary.org/obo/GO_0005623"}],"text":"Lentiviral shRNAs, Packaging and Transduction\npLKO.1-puro lentiviral vectors carrying short hairpin RNAs (shRNA) specific for human and mouse cebpb were used in these studies. Because the human and mouse cebpb 3′ untranslated regions are not identical, these species-specific shRNAs cannot knockdown expression of endogenous C/EBP-β when used on cells of the other species; therefore we used the human CEBPB shRNA as a control in these experiments as in [38]. To produce lentiviral particles, HEK-293T cells were transfected with a mixture of plasmids: each shRNA expression plasmid (1 µg), pCMV-ΔR8.2dvpr packaging plasmid (0.75 µg), and pCMV-VSV-G envelope plasmid (0.25 µg) using Lipofectamine 2000 reagent (Invitrogen). The lentiviral supernatant was collected 48 hrs after transfection, cleared by centrifugation at 2,000 g for 10 mins and passed through a 0.45 µm filter. The target cells were treated with the lentiviral supernatant and 8 µg/ml Polybrene (American Bioanalytical) for 24 hrs. The lentiviral supernatant was replaced with fresh growth media and incubated further for 72 hrs to allow for effective gene knockdown. C/EBP-β knockdown was confirmed by western blot analysis."}

sentences

{"project":"sentences","denotations":[{"id":"T6639","span":{"begin":1135,"end":1192},"obj":"Sentence"},{"id":"T6638","span":{"begin":999,"end":1134},"obj":"Sentence"},{"id":"T6637","span":{"begin":878,"end":998},"obj":"Sentence"},{"id":"T6636","span":{"begin":724,"end":877},"obj":"Sentence"},{"id":"T6635","span":{"begin":460,"end":723},"obj":"Sentence"},{"id":"T6634","span":{"begin":176,"end":459},"obj":"Sentence"},{"id":"T6633","span":{"begin":46,"end":175},"obj":"Sentence"},{"id":"T6632","span":{"begin":0,"end":45},"obj":"Sentence"},{"id":"T76","span":{"begin":0,"end":45},"obj":"Sentence"},{"id":"T77","span":{"begin":46,"end":175},"obj":"Sentence"},{"id":"T78","span":{"begin":176,"end":459},"obj":"Sentence"},{"id":"T79","span":{"begin":460,"end":723},"obj":"Sentence"},{"id":"T80","span":{"begin":724,"end":877},"obj":"Sentence"},{"id":"T81","span":{"begin":878,"end":998},"obj":"Sentence"},{"id":"T82","span":{"begin":999,"end":1134},"obj":"Sentence"},{"id":"T83","span":{"begin":1135,"end":1192},"obj":"Sentence"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/ontology/tao.owl#"}],"text":"Lentiviral shRNAs, Packaging and Transduction\npLKO.1-puro lentiviral vectors carrying short hairpin RNAs (shRNA) specific for human and mouse cebpb were used in these studies. Because the human and mouse cebpb 3′ untranslated regions are not identical, these species-specific shRNAs cannot knockdown expression of endogenous C/EBP-β when used on cells of the other species; therefore we used the human CEBPB shRNA as a control in these experiments as in [38]. To produce lentiviral particles, HEK-293T cells were transfected with a mixture of plasmids: each shRNA expression plasmid (1 µg), pCMV-ΔR8.2dvpr packaging plasmid (0.75 µg), and pCMV-VSV-G envelope plasmid (0.25 µg) using Lipofectamine 2000 reagent (Invitrogen). The lentiviral supernatant was collected 48 hrs after transfection, cleared by centrifugation at 2,000 g for 10 mins and passed through a 0.45 µm filter. The target cells were treated with the lentiviral supernatant and 8 µg/ml Polybrene (American Bioanalytical) for 24 hrs. The lentiviral supernatant was replaced with fresh growth media and incubated further for 72 hrs to allow for effective gene knockdown. C/EBP-β knockdown was confirmed by western blot analysis."}

simple1

{"project":"simple1","denotations":[{"id":"T6664","span":{"begin":1135,"end":1142},"obj":"Protein"},{"id":"T6663","span":{"begin":325,"end":332},"obj":"Protein"},{"id":"T6662","span":{"begin":204,"end":209},"obj":"Protein"},{"id":"T6661","span":{"begin":142,"end":147},"obj":"Protein"}],"text":"Lentiviral shRNAs, Packaging and Transduction\npLKO.1-puro lentiviral vectors carrying short hairpin RNAs (shRNA) specific for human and mouse cebpb were used in these studies. Because the human and mouse cebpb 3′ untranslated regions are not identical, these species-specific shRNAs cannot knockdown expression of endogenous C/EBP-β when used on cells of the other species; therefore we used the human CEBPB shRNA as a control in these experiments as in [38]. To produce lentiviral particles, HEK-293T cells were transfected with a mixture of plasmids: each shRNA expression plasmid (1 µg), pCMV-ΔR8.2dvpr packaging plasmid (0.75 µg), and pCMV-VSV-G envelope plasmid (0.25 µg) using Lipofectamine 2000 reagent (Invitrogen). The lentiviral supernatant was collected 48 hrs after transfection, cleared by centrifugation at 2,000 g for 10 mins and passed through a 0.45 µm filter. The target cells were treated with the lentiviral supernatant and 8 µg/ml Polybrene (American Bioanalytical) for 24 hrs. The lentiviral supernatant was replaced with fresh growth media and incubated further for 72 hrs to allow for effective gene knockdown. C/EBP-β knockdown was confirmed by western blot analysis."}

BioNLP16_DUT

{"project":"BioNLP16_DUT","denotations":[{"id":"T6870","span":{"begin":1143,"end":1152},"obj":"Negative_regulation"},{"id":"T6869","span":{"begin":300,"end":310},"obj":"Gene_expression"},{"id":"T6868","span":{"begin":290,"end":299},"obj":"Negative_regulation"},{"id":"T6867","span":{"begin":1135,"end":1142},"obj":"Protein"},{"id":"T6866","span":{"begin":325,"end":332},"obj":"Protein"},{"id":"T6865","span":{"begin":204,"end":209},"obj":"Protein"},{"id":"T6864","span":{"begin":142,"end":147},"obj":"Protein"}],"relations":[{"id":"R4950","pred":"themeOf","subj":"T6866","obj":"T6869"},{"id":"R4951","pred":"themeOf","subj":"T6867","obj":"T6870"},{"id":"R4952","pred":"themeOf","subj":"T6869","obj":"T6868"}],"text":"Lentiviral shRNAs, Packaging and Transduction\npLKO.1-puro lentiviral vectors carrying short hairpin RNAs (shRNA) specific for human and mouse cebpb were used in these studies. Because the human and mouse cebpb 3′ untranslated regions are not identical, these species-specific shRNAs cannot knockdown expression of endogenous C/EBP-β when used on cells of the other species; therefore we used the human CEBPB shRNA as a control in these experiments as in [38]. To produce lentiviral particles, HEK-293T cells were transfected with a mixture of plasmids: each shRNA expression plasmid (1 µg), pCMV-ΔR8.2dvpr packaging plasmid (0.75 µg), and pCMV-VSV-G envelope plasmid (0.25 µg) using Lipofectamine 2000 reagent (Invitrogen). The lentiviral supernatant was collected 48 hrs after transfection, cleared by centrifugation at 2,000 g for 10 mins and passed through a 0.45 µm filter. The target cells were treated with the lentiviral supernatant and 8 µg/ml Polybrene (American Bioanalytical) for 24 hrs. The lentiviral supernatant was replaced with fresh growth media and incubated further for 72 hrs to allow for effective gene knockdown. C/EBP-β knockdown was confirmed by western blot analysis."}

BioNLP16_Messiy

{"project":"BioNLP16_Messiy","denotations":[{"id":"T6893","span":{"begin":1143,"end":1152},"obj":"Negative_regulation"},{"id":"T6892","span":{"begin":300,"end":310},"obj":"Gene_expression"},{"id":"T6891","span":{"begin":290,"end":299},"obj":"Negative_regulation"},{"id":"T6890","span":{"begin":1135,"end":1142},"obj":"Protein"},{"id":"T6889","span":{"begin":325,"end":332},"obj":"Protein"},{"id":"T6888","span":{"begin":204,"end":209},"obj":"Protein"},{"id":"T6887","span":{"begin":142,"end":147},"obj":"Protein"}],"relations":[{"id":"R4957","pred":"themeOf","subj":"T6889","obj":"T6891"},{"id":"R4958","pred":"themeOf","subj":"T6889","obj":"T6892"},{"id":"R4959","pred":"themeOf","subj":"T6890","obj":"T6893"},{"id":"R4960","pred":"themeOf","subj":"T6892","obj":"T6891"}],"text":"Lentiviral shRNAs, Packaging and Transduction\npLKO.1-puro lentiviral vectors carrying short hairpin RNAs (shRNA) specific for human and mouse cebpb were used in these studies. Because the human and mouse cebpb 3′ untranslated regions are not identical, these species-specific shRNAs cannot knockdown expression of endogenous C/EBP-β when used on cells of the other species; therefore we used the human CEBPB shRNA as a control in these experiments as in [38]. To produce lentiviral particles, HEK-293T cells were transfected with a mixture of plasmids: each shRNA expression plasmid (1 µg), pCMV-ΔR8.2dvpr packaging plasmid (0.75 µg), and pCMV-VSV-G envelope plasmid (0.25 µg) using Lipofectamine 2000 reagent (Invitrogen). The lentiviral supernatant was collected 48 hrs after transfection, cleared by centrifugation at 2,000 g for 10 mins and passed through a 0.45 µm filter. The target cells were treated with the lentiviral supernatant and 8 µg/ml Polybrene (American Bioanalytical) for 24 hrs. The lentiviral supernatant was replaced with fresh growth media and incubated further for 72 hrs to allow for effective gene knockdown. C/EBP-β knockdown was confirmed by western blot analysis."}

DLUT931

{"project":"DLUT931","denotations":[{"id":"T6881","span":{"begin":1143,"end":1152},"obj":"Negative_regulation"},{"id":"T6880","span":{"begin":290,"end":299},"obj":"Negative_regulation"},{"id":"T6879","span":{"begin":300,"end":310},"obj":"Gene_expression"},{"id":"T6878","span":{"begin":1135,"end":1142},"obj":"Protein"},{"id":"T6877","span":{"begin":325,"end":332},"obj":"Protein"},{"id":"T6876","span":{"begin":204,"end":209},"obj":"Protein"},{"id":"T6875","span":{"begin":142,"end":147},"obj":"Protein"}],"relations":[{"id":"R4953","pred":"themeOf","subj":"T6877","obj":"T6879"},{"id":"R4954","pred":"themeOf","subj":"T6878","obj":"T6881"},{"id":"R4955","pred":"themeOf","subj":"T6879","obj":"T6880"}],"text":"Lentiviral shRNAs, Packaging and Transduction\npLKO.1-puro lentiviral vectors carrying short hairpin RNAs (shRNA) specific for human and mouse cebpb were used in these studies. Because the human and mouse cebpb 3′ untranslated regions are not identical, these species-specific shRNAs cannot knockdown expression of endogenous C/EBP-β when used on cells of the other species; therefore we used the human CEBPB shRNA as a control in these experiments as in [38]. To produce lentiviral particles, HEK-293T cells were transfected with a mixture of plasmids: each shRNA expression plasmid (1 µg), pCMV-ΔR8.2dvpr packaging plasmid (0.75 µg), and pCMV-VSV-G envelope plasmid (0.25 µg) using Lipofectamine 2000 reagent (Invitrogen). The lentiviral supernatant was collected 48 hrs after transfection, cleared by centrifugation at 2,000 g for 10 mins and passed through a 0.45 µm filter. The target cells were treated with the lentiviral supernatant and 8 µg/ml Polybrene (American Bioanalytical) for 24 hrs. The lentiviral supernatant was replaced with fresh growth media and incubated further for 72 hrs to allow for effective gene knockdown. C/EBP-β knockdown was confirmed by western blot analysis."}

bionlp-st-ge-2016-test-ihmc

{"project":"bionlp-st-ge-2016-test-ihmc","denotations":[{"id":"T6926","span":{"begin":1089,"end":1133},"obj":"Protein"},{"id":"T6925","span":{"begin":204,"end":209},"obj":"Protein"},{"id":"T6924","span":{"begin":739,"end":750},"obj":"Entity"},{"id":"T6923","span":{"begin":683,"end":722},"obj":"Entity"},{"id":"T6922","span":{"begin":1135,"end":1142},"obj":"Protein"},{"id":"T6921","span":{"begin":991,"end":997},"obj":"Protein"},{"id":"T6920","span":{"begin":765,"end":771},"obj":"Protein"},{"id":"T6919","span":{"begin":410,"end":413},"obj":"Protein"},{"id":"T6918","span":{"begin":346,"end":372},"obj":"Entity"},{"id":"T6917","span":{"begin":878,"end":894},"obj":"Entity"},{"id":"T6916","span":{"begin":1119,"end":1123},"obj":"Protein"},{"id":"T6915","span":{"begin":913,"end":939},"obj":"Entity"},{"id":"T6914","span":{"begin":471,"end":492},"obj":"Entity"},{"id":"T6913","span":{"begin":999,"end":1025},"obj":"Entity"},{"id":"T6912","span":{"begin":944,"end":986},"obj":"Entity"},{"id":"T6911","span":{"begin":402,"end":407},"obj":"Protein"},{"id":"T6910","span":{"begin":136,"end":147},"obj":"Protein"},{"id":"T6909","span":{"begin":525,"end":551},"obj":"Entity"},{"id":"T6908","span":{"begin":311,"end":330},"obj":"Protein"}],"text":"Lentiviral shRNAs, Packaging and Transduction\npLKO.1-puro lentiviral vectors carrying short hairpin RNAs (shRNA) specific for human and mouse cebpb were used in these studies. Because the human and mouse cebpb 3′ untranslated regions are not identical, these species-specific shRNAs cannot knockdown expression of endogenous C/EBP-β when used on cells of the other species; therefore we used the human CEBPB shRNA as a control in these experiments as in [38]. To produce lentiviral particles, HEK-293T cells were transfected with a mixture of plasmids: each shRNA expression plasmid (1 µg), pCMV-ΔR8.2dvpr packaging plasmid (0.75 µg), and pCMV-VSV-G envelope plasmid (0.25 µg) using Lipofectamine 2000 reagent (Invitrogen). The lentiviral supernatant was collected 48 hrs after transfection, cleared by centrifugation at 2,000 g for 10 mins and passed through a 0.45 µm filter. The target cells were treated with the lentiviral supernatant and 8 µg/ml Polybrene (American Bioanalytical) for 24 hrs. The lentiviral supernatant was replaced with fresh growth media and incubated further for 72 hrs to allow for effective gene knockdown. C/EBP-β knockdown was confirmed by western blot analysis."}

bionlp-st-ge-2016-test-tees

{"project":"bionlp-st-ge-2016-test-tees","denotations":[{"id":"T6900","span":{"begin":1143,"end":1152},"obj":"Negative_regulation"},{"id":"T6899","span":{"begin":1135,"end":1142},"obj":"Protein"},{"id":"T6898","span":{"begin":290,"end":299},"obj":"Negative_regulation"},{"id":"T6897","span":{"begin":419,"end":426},"obj":"Regulation"},{"id":"T6896","span":{"begin":300,"end":310},"obj":"Gene_expression"},{"id":"T6895","span":{"begin":396,"end":413},"obj":"Protein"},{"id":"T6894","span":{"begin":325,"end":332},"obj":"Protein"}],"relations":[{"id":"R4961","pred":"themeOf","subj":"T6894","obj":"T6896"},{"id":"R4962","pred":"themeOf","subj":"T6896","obj":"T6897"},{"id":"R4963","pred":"themeOf","subj":"T6896","obj":"T6898"},{"id":"R4964","pred":"themeOf","subj":"T6899","obj":"T6900"}],"text":"Lentiviral shRNAs, Packaging and Transduction\npLKO.1-puro lentiviral vectors carrying short hairpin RNAs (shRNA) specific for human and mouse cebpb were used in these studies. Because the human and mouse cebpb 3′ untranslated regions are not identical, these species-specific shRNAs cannot knockdown expression of endogenous C/EBP-β when used on cells of the other species; therefore we used the human CEBPB shRNA as a control in these experiments as in [38]. To produce lentiviral particles, HEK-293T cells were transfected with a mixture of plasmids: each shRNA expression plasmid (1 µg), pCMV-ΔR8.2dvpr packaging plasmid (0.75 µg), and pCMV-VSV-G envelope plasmid (0.25 µg) using Lipofectamine 2000 reagent (Invitrogen). The lentiviral supernatant was collected 48 hrs after transfection, cleared by centrifugation at 2,000 g for 10 mins and passed through a 0.45 µm filter. The target cells were treated with the lentiviral supernatant and 8 µg/ml Polybrene (American Bioanalytical) for 24 hrs. The lentiviral supernatant was replaced with fresh growth media and incubated further for 72 hrs to allow for effective gene knockdown. C/EBP-β knockdown was confirmed by western blot analysis."}

testone

{"project":"testone","denotations":[{"id":"T6620","span":{"begin":1143,"end":1152},"obj":"Negative_regulation"},{"id":"T6619","span":{"begin":463,"end":470},"obj":"Gene_expression"},{"id":"T6618","span":{"begin":300,"end":310},"obj":"Gene_expression"},{"id":"T6617","span":{"begin":290,"end":299},"obj":"Negative_regulation"},{"id":"T6616","span":{"begin":1135,"end":1142},"obj":"Protein"},{"id":"T6615","span":{"begin":325,"end":332},"obj":"Protein"},{"id":"T6614","span":{"begin":204,"end":209},"obj":"Protein"},{"id":"T6613","span":{"begin":142,"end":147},"obj":"Protein"}],"relations":[{"id":"R4736","pred":"themeOf","subj":"T6615","obj":"T6618"},{"id":"R4737","pred":"themeOf","subj":"T6616","obj":"T6620"}],"text":"Lentiviral shRNAs, Packaging and Transduction\npLKO.1-puro lentiviral vectors carrying short hairpin RNAs (shRNA) specific for human and mouse cebpb were used in these studies. Because the human and mouse cebpb 3′ untranslated regions are not identical, these species-specific shRNAs cannot knockdown expression of endogenous C/EBP-β when used on cells of the other species; therefore we used the human CEBPB shRNA as a control in these experiments as in [38]. To produce lentiviral particles, HEK-293T cells were transfected with a mixture of plasmids: each shRNA expression plasmid (1 µg), pCMV-ΔR8.2dvpr packaging plasmid (0.75 µg), and pCMV-VSV-G envelope plasmid (0.25 µg) using Lipofectamine 2000 reagent (Invitrogen). The lentiviral supernatant was collected 48 hrs after transfection, cleared by centrifugation at 2,000 g for 10 mins and passed through a 0.45 µm filter. The target cells were treated with the lentiviral supernatant and 8 µg/ml Polybrene (American Bioanalytical) for 24 hrs. The lentiviral supernatant was replaced with fresh growth media and incubated further for 72 hrs to allow for effective gene knockdown. C/EBP-β knockdown was confirmed by western blot analysis."}

test3

{"project":"test3","denotations":[{"id":"T6631","span":{"begin":1143,"end":1152},"obj":"Negative_regulation"},{"id":"T6630","span":{"begin":1135,"end":1142},"obj":"Protein"},{"id":"T6629","span":{"begin":325,"end":332},"obj":"Protein"},{"id":"T6628","span":{"begin":300,"end":310},"obj":"Gene_expression"},{"id":"T6627","span":{"begin":290,"end":299},"obj":"Negative_regulation"},{"id":"T6626","span":{"begin":204,"end":209},"obj":"Protein"},{"id":"T6625","span":{"begin":142,"end":147},"obj":"Protein"},{"id":"T6624","span":{"begin":1135,"end":1142},"obj":"Protein"},{"id":"T6623","span":{"begin":325,"end":332},"obj":"Protein"},{"id":"T6622","span":{"begin":204,"end":209},"obj":"Protein"},{"id":"T6621","span":{"begin":142,"end":147},"obj":"Protein"}],"relations":[{"id":"R4738","pred":"causeOf","subj":"T6626","obj":"T6627"},{"id":"R4739","pred":"themeOf","subj":"T6628","obj":"T6627"},{"id":"R4740","pred":"themeOf","subj":"T6629","obj":"T6628"},{"id":"R4741","pred":"themeOf","subj":"T6630","obj":"T6631"}],"text":"Lentiviral shRNAs, Packaging and Transduction\npLKO.1-puro lentiviral vectors carrying short hairpin RNAs (shRNA) specific for human and mouse cebpb were used in these studies. Because the human and mouse cebpb 3′ untranslated regions are not identical, these species-specific shRNAs cannot knockdown expression of endogenous C/EBP-β when used on cells of the other species; therefore we used the human CEBPB shRNA as a control in these experiments as in [38]. To produce lentiviral particles, HEK-293T cells were transfected with a mixture of plasmids: each shRNA expression plasmid (1 µg), pCMV-ΔR8.2dvpr packaging plasmid (0.75 µg), and pCMV-VSV-G envelope plasmid (0.25 µg) using Lipofectamine 2000 reagent (Invitrogen). The lentiviral supernatant was collected 48 hrs after transfection, cleared by centrifugation at 2,000 g for 10 mins and passed through a 0.45 µm filter. The target cells were treated with the lentiviral supernatant and 8 µg/ml Polybrene (American Bioanalytical) for 24 hrs. The lentiviral supernatant was replaced with fresh growth media and incubated further for 72 hrs to allow for effective gene knockdown. C/EBP-β knockdown was confirmed by western blot analysis."}

PMC:3589482 / 12598-13790 JSONTXT

Annnotations TAB JSON ListView MergeView

PMC:3589482 / 12598-13790 JSON TXT