Effect of CPNE5 on transcriptional activity of NF-κB
Yan Wu
Abstract
Yan Wu
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.
A significance statement is not available in the OpenAlex record.
A contribution statement is not available in the OpenAlex record.
Method details are not available in the OpenAlex metadata.
Findings are not separately available in the OpenAlex metadata.
Limitations are not available in the OpenAlex metadata.
Application details are not available in the OpenAlex metadata.
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