PMC:3245220 / 14033-15167 JSONTXT

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Figure 3 Inhibition of PKC or NF-κB induces apoptosis in MDMs.\n(A) MDMs were pre-incubated in RPMI medium containing inhibitors (Ro-31-8220: 5 µM, Gö-6976: 5 µM, BAPTA/AM: 2.5 µM) for 30 minutes prior to the addition of M-CSF. 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Figure 3 Inhibition of PKC or NF-κB induces apoptosis in MDMs.\n(A) MDMs were pre-incubated in RPMI medium containing inhibitors (Ro-31-8220: 5 µM, Gö-6976: 5 µM, BAPTA/AM: 2.5 µM) for 30 minutes prior to the addition of M-CSF. As a control, untreated cells were incubated with dimethyl sulfoxide (DMSO). Cell lysates were resolved by SDS-PAGE and immunoblotted with antibody recognizing the active cleaved form of caspase-3. The blots were reblotted with β-actin that served as a loading control. The ratio of active caspase-3 bands (17 kD and 19 kD) to β-actin control was determined by densitometry analysis (bottom panel). Data represents the mean ± S.E.M from two independent donors. (B) Apoptosis of the treated MDMs was also measured by Annexin V-FITC and propidium iodine (PI) staining and analyzed by flow cytometry. The percentage of surviving cells (Annexin V/PI negative) for the cells treated with vehicle/M-CSF was arbitrarily set as 100. Data shown represent the mean ± S.E.M from three independent experiments. *The p-values of inhibitors/M-CSF compared to vehicle/M-CSF were ≤0.05. "}

{"project":"test2","denotations":[{"id":"T23538","span":{"begin":255,"end":260},"obj":"Protein"},{"id":"T23539","span":{"begin":490,"end":497},"obj":"Protein"},{"id":"T23540","span":{"begin":552,"end":561},"obj":"Protein"},{"id":"T23541","span":{"begin":589,"end":596},"obj":"Protein"},{"id":"T23542","span":{"begin":778,"end":787},"obj":"Protein"},{"id":"T23543","span":{"begin":895,"end":904},"obj":"Protein"},{"id":"T23544","span":{"begin":908,"end":916},"obj":"Negative_regulation"},{"id":"T23545","span":{"begin":953,"end":958},"obj":"Protein"},{"id":"T23546","span":{"begin":1089,"end":1094},"obj":"Protein"},{"id":"T23547","span":{"begin":1115,"end":1120},"obj":"Protein"}],"relations":[{"id":"R17950","pred":"themeOf","subj":"T23543","obj":"T23544"}],"text":"10.1371/journal.pone.0028081.g003 Figure 3 Inhibition of PKC or NF-κB induces apoptosis in MDMs.\n(A) MDMs were pre-incubated in RPMI medium containing inhibitors (Ro-31-8220: 5 µM, Gö-6976: 5 µM, BAPTA/AM: 2.5 µM) for 30 minutes prior to the addition of M-CSF. As a control, untreated cells were incubated with dimethyl sulfoxide (DMSO). Cell lysates were resolved by SDS-PAGE and immunoblotted with antibody recognizing the active cleaved form of caspase-3. The blots were reblotted with β-actin that served as a loading control. The ratio of active caspase-3 bands (17 kD and 19 kD) to β-actin control was determined by densitometry analysis (bottom panel). Data represents the mean ± S.E.M from two independent donors. (B) Apoptosis of the treated MDMs was also measured by Annexin V-FITC and propidium iodine (PI) staining and analyzed by flow cytometry. The percentage of surviving cells (Annexin V/PI negative) for the cells treated with vehicle/M-CSF was arbitrarily set as 100. Data shown represent the mean ± S.E.M from three independent experiments. *The p-values of inhibitors/M-CSF compared to vehicle/M-CSF were ≤0.05. "}