PMC:3245220 / 18292-19576 JSONTXT

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{"project":"GO-MF","denotations":[{"id":"T24953","span":{"begin":58,"end":70},"obj":"http://purl.obolibrary.org/obo/GO_0004697"},{"id":"T24954","span":{"begin":391,"end":399},"obj":"http://purl.obolibrary.org/obo/GO_0003823"},{"id":"T24955","span":{"begin":709,"end":719},"obj":"http://purl.obolibrary.org/obo/GO_0003823"},{"id":"T24956","span":{"begin":961,"end":971},"obj":"http://purl.obolibrary.org/obo/GO_0003823"}],"text":"10.1371/journal.pone.0028081.g005 Figure 5 M-CSF-induced PKC activity does not regulate IκBα degradation but regulates the phosphorylation of NF-κB p65 at Ser276.\n(A) MDMs were pretreated with cycloheximide (CHX) in the absence or presence of Ro-31-8220 for 30 minutes prior to M-CSF stimulation for the indicated times. Whole cell lysates were subjected to Western blotting with anti-IκBα antibody. Data are representative of three independent experiments. (B) MDMs were pretreated with either vehicle or Ro-31-8220 for 30 minutes before the addition of M-CSF for 10 minutes. Whole cell lysates were resolved by SDS-PAGE and phospho-Ser276 or phospho-Ser536 of NF-κB p65 was detected using phospho-specific antibodies to either residue of NF-κB p65. (C) Whole cell lysates of RAW 264.7 cells treated with vehicle control or Ro-31-8220 in the absence or presence of M-CSF were subjected to Western blot analysis with phospho-Ser276 or phospho-Ser536 NF-κB p65 antibodies. (D) Cytosolic and nuclear fractions of RAW 264.7 were obtained from the treated cells and immunoblotted for phospho-NF-κB. The purity of the cytosolic and nuclear fractions was confirmed by immunoblotting with GAPDH and Lamin B, respectively. Shown are representative blots from three independent experiments.\n\n"}

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Figure 5 M-CSF-induced PKC activity does not regulate IκBα degradation but regulates the phosphorylation of NF-κB p65 at Ser276.\n(A) MDMs were pretreated with cycloheximide (CHX) in the absence or presence of Ro-31-8220 for 30 minutes prior to M-CSF stimulation for the indicated times. Whole cell lysates were subjected to Western blotting with anti-IκBα antibody. Data are representative of three independent experiments. (B) MDMs were pretreated with either vehicle or Ro-31-8220 for 30 minutes before the addition of M-CSF for 10 minutes. Whole cell lysates were resolved by SDS-PAGE and phospho-Ser276 or phospho-Ser536 of NF-κB p65 was detected using phospho-specific antibodies to either residue of NF-κB p65. (C) Whole cell lysates of RAW 264.7 cells treated with vehicle control or Ro-31-8220 in the absence or presence of M-CSF were subjected to Western blot analysis with phospho-Ser276 or phospho-Ser536 NF-κB p65 antibodies. (D) Cytosolic and nuclear fractions of RAW 264.7 were obtained from the treated cells and immunoblotted for phospho-NF-κB. The purity of the cytosolic and nuclear fractions was confirmed by immunoblotting with GAPDH and Lamin B, respectively. Shown are representative blots from three independent experiments.\n\n"}

{"project":"test2","denotations":[{"id":"T24487","span":{"begin":44,"end":49},"obj":"Protein"},{"id":"T24488","span":{"begin":80,"end":88},"obj":"Regulation"},{"id":"T24489","span":{"begin":89,"end":93},"obj":"Protein"},{"id":"T24490","span":{"begin":94,"end":105},"obj":"Protein_catabolism"},{"id":"T24491","span":{"begin":110,"end":119},"obj":"Regulation"},{"id":"T24492","span":{"begin":124,"end":139},"obj":"Phosphorylation"},{"id":"T24493","span":{"begin":149,"end":152},"obj":"Protein"},{"id":"T24494","span":{"begin":156,"end":162},"obj":"Entity"},{"id":"T24495","span":{"begin":279,"end":284},"obj":"Protein"},{"id":"T24496","span":{"begin":556,"end":561},"obj":"Protein"},{"id":"T24497","span":{"begin":645,"end":652},"obj":"Phosphorylation"},{"id":"T24498","span":{"begin":669,"end":672},"obj":"Protein"},{"id":"T24499","span":{"begin":747,"end":750},"obj":"Protein"},{"id":"T24500","span":{"begin":867,"end":872},"obj":"Protein"},{"id":"T24501","span":{"begin":918,"end":925},"obj":"Phosphorylation"},{"id":"T24502","span":{"begin":936,"end":943},"obj":"Phosphorylation"}],"relations":[{"id":"R18669","pred":"causeOf","subj":"T24487","obj":"T24488"},{"id":"R18670","pred":"causeOf","subj":"T24487","obj":"T24491"},{"id":"R18671","pred":"themeOf","subj":"T24489","obj":"T24490"},{"id":"R18672","pred":"themeOf","subj":"T24490","obj":"T24488"},{"id":"R18673","pred":"themeOf","subj":"T24492","obj":"T24491"},{"id":"R18674","pred":"partOf","subj":"T24494","obj":"T24493"},{"id":"R18675","pred":"themeOf","subj":"T24494","obj":"T24492"}],"attributes":[{"id":"M257","pred":"Negation","subj":"T24487","obj":"true"},{"id":"M259","pred":"Speculation","subj":"T24492","obj":"true"},{"id":"M258","pred":"Speculation","subj":"T24491","obj":"true"}],"text":"10.1371/journal.pone.0028081.g005 Figure 5 M-CSF-induced PKC activity does not regulate IκBα degradation but regulates the phosphorylation of NF-κB p65 at Ser276.\n(A) MDMs were pretreated with cycloheximide (CHX) in the absence or presence of Ro-31-8220 for 30 minutes prior to M-CSF stimulation for the indicated times. Whole cell lysates were subjected to Western blotting with anti-IκBα antibody. Data are representative of three independent experiments. (B) MDMs were pretreated with either vehicle or Ro-31-8220 for 30 minutes before the addition of M-CSF for 10 minutes. Whole cell lysates were resolved by SDS-PAGE and phospho-Ser276 or phospho-Ser536 of NF-κB p65 was detected using phospho-specific antibodies to either residue of NF-κB p65. (C) Whole cell lysates of RAW 264.7 cells treated with vehicle control or Ro-31-8220 in the absence or presence of M-CSF were subjected to Western blot analysis with phospho-Ser276 or phospho-Ser536 NF-κB p65 antibodies. (D) Cytosolic and nuclear fractions of RAW 264.7 were obtained from the treated cells and immunoblotted for phospho-NF-κB. The purity of the cytosolic and nuclear fractions was confirmed by immunoblotting with GAPDH and Lamin B, respectively. Shown are representative blots from three independent experiments.\n\n"}