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untreated or treated for 24 h with 0.5 μg/ml adriamycin, were lysed on the dish in a buffer consisting of 50 mM Tris (pH 8.0), 5 mM EDTA, 150 mM NaCl, 0.5% Nonidet P-40, 1 mM PMSF, 1 mM sodium vanadate, 10 mM NaF and Complete Mini Protease Inhibitors (Roche Diagnostics) at 4°C for 30 min. Lysates were scraped, then spun at 6000× g at 4°C for 10 min. Protein concentration in the supernatant was determined using the Bio-Rad DC protein assay. Lysates were separated on single percentage SDS/PAGE gels, then electrophoretically transferred to poly(vinylidene difluoride), using standard procedures.Blots were incubated in 5% non-fat dried milk in TBST (0.02 M Tris, pH 7.6/0.35 M NaCl/0.1% Tween-20) for 1 h at room temperature before probing with primary antibodies against p53 (CM-5, Novacastra) and -actin (Sigma). Secondary antibodies used include peroxidase-conjugated goat anti-mouse IgG and anti-rabbit IgG (Pierce). Probed blots were incubated with Pierce Supersignal West Pico chemiluminescent substrate and exposed to X-ray films."}

{"project":"craft-ca-core-ex-dev","denotations":[{"id":"T8038","span":{"begin":0,"end":5},"obj":"CL_GO_EXT:cell"},{"id":"T8039","span":{"begin":52,"end":62},"obj":"CHEBI:28748"},{"id":"T8040","span":{"begin":69,"end":74},"obj":"GO:0019835"},{"id":"T8041","span":{"begin":92,"end":98},"obj":"CHEBI_CHMO_EXT:buffer_solution"},{"id":"T8042","span":{"begin":119,"end":123},"obj":"CHEBI:9754"},{"id":"T8043","span":{"begin":139,"end":143},"obj":"CHEBI_EXT:EDTA"},{"id":"T8044","span":{"begin":152,"end":156},"obj":"CHEBI:26710"},{"id":"T8045","span":{"begin":163,"end":175},"obj":"CHEBI:78708"},{"id":"T8046","span":{"begin":182,"end":186},"obj":"CHEBI:8102"},{"id":"T8047","span":{"begin":193,"end":208},"obj":"CHEBI:35607"},{"id":"T8048","span":{"begin":216,"end":219},"obj":"CHEBI:28741"},{"id":"T8049","span":{"begin":238,"end":257},"obj":"CHEBI_GO_EXT:peptidase_or_protease_or_proteinase_inhibitor"},{"id":"T8050","span":{"begin":359,"end":366},"obj":"CHEBI_PR_EXT:protein"},{"id":"T8051","span":{"begin":436,"end":443},"obj":"CHEBI_PR_EXT:protein"},{"id":"T8052","span":{"begin":495,"end":498},"obj":"CHEBI:8984"},{"id":"T8053","span":{"begin":535,"end":546},"obj":"GO_EXT:biological_movement_or_translocation_process"},{"id":"T8054","span":{"begin":550,"end":576},"obj":"CHEBI:53250"},{"id":"T8055","span":{"begin":636,"end":639},"obj":"CHEBI_UBERON_EXT:triglyceride_or_adipose_tissue"},{"id":"T8056","span":{"begin":646,"end":650},"obj":"UBERON:0001913"},{"id":"T8057","span":{"begin":667,"end":671},"obj":"CHEBI:9754"},{"id":"T8058","span":{"begin":687,"end":691},"obj":"CHEBI:26710"},{"id":"T8059","span":{"begin":697,"end":705},"obj":"CHEBI:53424"},{"id":"T8060","span":{"begin":763,"end":773},"obj":"GO:0042571"},{"id":"T8061","span":{"begin":782,"end":785},"obj":"PR_EXT:000003035"},{"id":"T8062","span":{"begin":835,"end":845},"obj":"GO:0042571"},{"id":"T8063","span":{"begin":859,"end":869},"obj":"GO_EXT:0004601"},{"id":"T8064","span":{"begin":870,"end":880},"obj":"MOP:0000779"},{"id":"T8065","span":{"begin":881,"end":885},"obj":"NCBITaxon:9925"},{"id":"T8066","span":{"begin":891,"end":896},"obj":"NCBITaxon:10088"},{"id":"T8067","span":{"begin":897,"end":900},"obj":"GO:0071735"},{"id":"T8068","span":{"begin":910,"end":916},"obj":"NCBITaxon:9986"},{"id":"T8069","span":{"begin":917,"end":920},"obj":"GO:0071735"}],"text":"Cells, untreated or treated for 24 h with 0.5 μg/ml adriamycin, were lysed on the dish in a buffer consisting of 50 mM Tris (pH 8.0), 5 mM EDTA, 150 mM NaCl, 0.5% Nonidet P-40, 1 mM PMSF, 1 mM sodium vanadate, 10 mM NaF and Complete Mini Protease Inhibitors (Roche Diagnostics) at 4°C for 30 min. Lysates were scraped, then spun at 6000× g at 4°C for 10 min. Protein concentration in the supernatant was determined using the Bio-Rad DC protein assay. Lysates were separated on single percentage SDS/PAGE gels, then electrophoretically transferred to poly(vinylidene difluoride), using standard procedures.Blots were incubated in 5% non-fat dried milk in TBST (0.02 M Tris, pH 7.6/0.35 M NaCl/0.1% Tween-20) for 1 h at room temperature before probing with primary antibodies against p53 (CM-5, Novacastra) and -actin (Sigma). Secondary antibodies used include peroxidase-conjugated goat anti-mouse IgG and anti-rabbit IgG (Pierce). Probed blots were incubated with Pierce Supersignal West Pico chemiluminescent substrate and exposed to X-ray films."}

{"project":"craft-ca-core-dev","denotations":[{"id":"T8015","span":{"begin":52,"end":62},"obj":"CHEBI:28748"},{"id":"T8016","span":{"begin":69,"end":74},"obj":"GO:0019835"},{"id":"T8017","span":{"begin":119,"end":123},"obj":"CHEBI:9754"},{"id":"T8018","span":{"begin":152,"end":156},"obj":"CHEBI:26710"},{"id":"T8019","span":{"begin":163,"end":175},"obj":"CHEBI:78708"},{"id":"T8020","span":{"begin":182,"end":186},"obj":"CHEBI:8102"},{"id":"T8021","span":{"begin":193,"end":208},"obj":"CHEBI:35607"},{"id":"T8022","span":{"begin":216,"end":219},"obj":"CHEBI:28741"},{"id":"T8023","span":{"begin":495,"end":498},"obj":"CHEBI:8984"},{"id":"T8024","span":{"begin":550,"end":576},"obj":"CHEBI:53250"},{"id":"T8025","span":{"begin":646,"end":650},"obj":"UBERON:0001913"},{"id":"T8026","span":{"begin":667,"end":671},"obj":"CHEBI:9754"},{"id":"T8027","span":{"begin":687,"end":691},"obj":"CHEBI:26710"},{"id":"T8028","span":{"begin":697,"end":705},"obj":"CHEBI:53424"},{"id":"T8029","span":{"begin":763,"end":773},"obj":"GO:0042571"},{"id":"T8030","span":{"begin":782,"end":785},"obj":"PR:000003035"},{"id":"T8031","span":{"begin":835,"end":845},"obj":"GO:0042571"},{"id":"T8032","span":{"begin":870,"end":880},"obj":"MOP:0000779"},{"id":"T8033","span":{"begin":881,"end":885},"obj":"NCBITaxon:9925"},{"id":"T8034","span":{"begin":891,"end":896},"obj":"NCBITaxon:10088"},{"id":"T8035","span":{"begin":897,"end":900},"obj":"GO:0071735"},{"id":"T8036","span":{"begin":910,"end":916},"obj":"NCBITaxon:9986"},{"id":"T8037","span":{"begin":917,"end":920},"obj":"GO:0071735"}],"text":"Cells, untreated or treated for 24 h with 0.5 μg/ml adriamycin, were lysed on the dish in a buffer consisting of 50 mM Tris (pH 8.0), 5 mM EDTA, 150 mM NaCl, 0.5% Nonidet P-40, 1 mM PMSF, 1 mM sodium vanadate, 10 mM NaF and Complete Mini Protease Inhibitors (Roche Diagnostics) at 4°C for 30 min. Lysates were scraped, then spun at 6000× g at 4°C for 10 min. Protein concentration in the supernatant was determined using the Bio-Rad DC protein assay. Lysates were separated on single percentage SDS/PAGE gels, then electrophoretically transferred to poly(vinylidene difluoride), using standard procedures.Blots were incubated in 5% non-fat dried milk in TBST (0.02 M Tris, pH 7.6/0.35 M NaCl/0.1% Tween-20) for 1 h at room temperature before probing with primary antibodies against p53 (CM-5, Novacastra) and -actin (Sigma). Secondary antibodies used include peroxidase-conjugated goat anti-mouse IgG and anti-rabbit IgG (Pierce). Probed blots were incubated with Pierce Supersignal West Pico chemiluminescent substrate and exposed to X-ray films."}