Mipu1 Protects H9c2 Myogenic Cells from Hydrogen Peroxide-Induced Apoptosis through Inhibition of the Expression of the Death Receptor Fas
Abstract
:1. Introduction
2. Results
2.1. RT-PCR, Quantitative Real-Time RT-PCR and Western Blotting Showed Expression of Mipu1 in H9c2 Myogenic Cells after Exposure to H2O2
2.2. Effects of Mipu1 Over-Expression on Apoptosis Induced by H2O2
2.3. Effects of Mipu1 Inhibition by RNAi on Apoptosis Induced by H2O2
2.4. Effects of Mipu1 Over-Expression on the Expression of Fas
2.5. Effects of Mipu1 Inhibition by RNAi on the Expression of Fas
2.6. Binding of Mipu1 to the Fas Promoter
2.7. Inhibition of the Luciferase Reporter Activity of Fas Promoter by Mipu1
3. Discussion
4. Methods
4.1. Cell Culture and Reagent
4.2. Plasmids Construction
4.3. RNA Extraction and Reverse-Transcription PCR (RT-PCR)
Genes | Primers |
---|---|
Mipu1 | Sence 5'-ATGCCTGCAGCCCGAGGGAAATC-3' |
Antisence 5'-CGATGATATTTGGCCTCCGGCAGGC-3' | |
GAPDH | Sence 5'-AACACAGTCCATGCCATCAC-3' |
Antisence 5'-TCCACCACCCTGTTGCTGTA-3' |
4.4. Quantitative Real-Time RT-PCR
4.5. Western Blotting Analysis
4.6. Gene Transfection
4.7. Quantification of Apoptotic Cells by Flow Cytometry
4.8. Chromatin Immunoprecipitation
4.9. Luciferase Reporter Gene Assay
4.10. Statistical Analysis
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Wang, G.; Jiang, L.; Song, J.; Zhou, S.-F.; Zhang, H.; Wang, K.; Xiao, X. Mipu1 Protects H9c2 Myogenic Cells from Hydrogen Peroxide-Induced Apoptosis through Inhibition of the Expression of the Death Receptor Fas. Int. J. Mol. Sci. 2014, 15, 18206-18220. https://doi.org/10.3390/ijms151018206
Wang G, Jiang L, Song J, Zhou S-F, Zhang H, Wang K, Xiao X. Mipu1 Protects H9c2 Myogenic Cells from Hydrogen Peroxide-Induced Apoptosis through Inhibition of the Expression of the Death Receptor Fas. International Journal of Molecular Sciences. 2014; 15(10):18206-18220. https://doi.org/10.3390/ijms151018206
Chicago/Turabian StyleWang, Guiliang, Lei Jiang, Juan Song, Shu-Feng Zhou, Huali Zhang, Kangkai Wang, and Xianzhong Xiao. 2014. "Mipu1 Protects H9c2 Myogenic Cells from Hydrogen Peroxide-Induced Apoptosis through Inhibition of the Expression of the Death Receptor Fas" International Journal of Molecular Sciences 15, no. 10: 18206-18220. https://doi.org/10.3390/ijms151018206