2009Junshi Yixue Kexueyuan yuankanRequires access

Effect of CPNE5 on transcriptional activity of NF-κB

Yan Wu

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Abstract

Objective:To study the effect and the mechanism of CPNE5 on the transcriptional activity of NF-κB induced by TNF-α.Methods:HEK293 cells transiently transfected with pcDNA3.1 or pcDNA3.1-CPNE5 were treated with TNF-α.Dual-Luciferase Reporter Assay was used to study the transcriptional activity of NF-κB among different groups.Effects of CPNE5 on nuclear translocation of NF-κB were exemined by Western blotting and immunofluorescence staining.Effects of CPNE5 on DNA binding activity of NF-κB were studied by electrophoretic mobility shift Assay(EMSA).Results:CPNE5 inhibited transactivation activity of NF-κB induced by TNF-α(P0.0001).Although CPNE5 had no effect on nuclear translocation of NF-κB,CPNE5 inhibited DNA binding activity of NF-κB induced by TNF-α.Conclusion:CPNE5 represses transactivation activity of NF-κB by inhibiting its DNA binding activity.

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Objective:To study the effect and the mechanism of CPNE5 on the transcriptional activity of NF-κB induced by TNF-α.Methods:HEK293 cells transiently transfected with pcDNA3.1 or pcDNA3.1-CPNE5 were treated with TNF-α.Dual-Luciferase Reporter Assay was used to study the transcriptional activity of NF-κB among different groups.Effects of CPNE5 on nuclear translocation of NF-κB were exemined by Western blotting and immunofluorescence staining.Effects of CPNE5 on DNA binding activity of NF-κB were studied by electrophoretic mobility shift Assay(EMSA).Results:CPNE5 inhibited transactivation activity of NF-κB induced by TNF-α(P0.0001).Although CPNE5 had no effect on nuclear translocation of NF-κB,CPNE5 inhibited DNA binding activity of NF-κB induced by TNF-α.Conclusion:CPNE5 represses transactivation activity of NF-κB by inhibiting its DNA binding activity.

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Available abstract

Objective:To study the effect and the mechanism of CPNE5 on the transcriptional activity of NF-κB induced by TNF-α.Methods:HEK293 cells transiently transfected with pcDNA3.1 or pcDNA3.1-CPNE5 were treated with TNF-α.Dual-Luciferase Reporter Assay was used to study the transcriptional activity of NF-κB among different groups.Effects of CPNE5 on nuclear translocation of NF-κB were exemined by Western blotting and immunofluorescence staining.Effects of CPNE5 on DNA binding activity of NF-κB were studied by electrophoretic mobility shift Assay(EMSA).Results:CPNE5 inhibited transactivation activity of NF-κB induced by TNF-α(P0.0001).Although CPNE5 had no effect on nuclear translocation of NF-κB,CPNE5 inhibited DNA binding activity of NF-κB induced by TNF-α.Conclusion:CPNE5 represses transactivation activity of NF-κB by inhibiting its DNA binding activity.

Key concepts: Transactivation, Electrophoretic mobility shift assay, NF-κB, Transfection, Molecular biology, Chemistry, Luciferase, Blot

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