BACE1 Inhibition Increases Susceptibility to Oxidative Stress by Promoting Mitochondrial Damage
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture
2.2. Transient Knockdown of BACE1 and BACE2
2.3. Oxidative Stress Protocol
2.4. Cytotoxicity Assay
2.5. β-Secretase Activity Assay
2.6. Amyloid β 40 Assay
2.7. DNA Fragmentation Detection
2.8. Mitochondrial Membrane Potential (Δψm)
2.9. Mitochondrial Morphology
2.10. Mitophagy
2.11. Mitochondria Isolation
2.12. Western Blot
2.13. Immunocytochemistry
2.14. Statistical Analysis
2.15. Data Resources
3. Results
3.1. Reduction of BACE1, but Not BACE2, Increases Susceptibility to Oxidative Stress
3.2. Effect of Oxidative Stress on BACE Expression and Aβ Secretion
3.3. Reduction in BACE1 Expression Results in Loss of Mitochondrial Membrane Potential
3.4. Reduction in BACE1 Expression Enhances the Induction of Mitochondrial Fission in Cells under Oxidative Stress
3.5. Mitophagic Machinery Is Upregulated in BACE1 Knockdown Cells under Oxidative Stress
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Francelin, C.; Mitter, S.K.; Qian, Q.; Barodia, S.K.; Ip, C.; Qi, X.; Gu, H.; Quigley, J.; Goldberg, M.S.; Grant, M.B.; et al. BACE1 Inhibition Increases Susceptibility to Oxidative Stress by Promoting Mitochondrial Damage. Antioxidants 2021, 10, 1539. https://doi.org/10.3390/antiox10101539
Francelin C, Mitter SK, Qian Q, Barodia SK, Ip C, Qi X, Gu H, Quigley J, Goldberg MS, Grant MB, et al. BACE1 Inhibition Increases Susceptibility to Oxidative Stress by Promoting Mitochondrial Damage. Antioxidants. 2021; 10(10):1539. https://doi.org/10.3390/antiox10101539
Chicago/Turabian StyleFrancelin, Carolina, Sayak K. Mitter, Qingwen Qian, Sandeep Kumar Barodia, Colin Ip, Xiaoping Qi, Hongmei Gu, Judith Quigley, Matthew S. Goldberg, Maria B. Grant, and et al. 2021. "BACE1 Inhibition Increases Susceptibility to Oxidative Stress by Promoting Mitochondrial Damage" Antioxidants 10, no. 10: 1539. https://doi.org/10.3390/antiox10101539