Mitochondrial Damage Induced by T-2 Mycotoxin on Human Skin—Fibroblast Hs68 Cell Line
Abstract
:1. Introduction
2. Results
2.1. Assessment of Mitochondrial Membrane Potential (MMP)
2.2. Assessment of Intracellular Reactive Oxygen Species (ROS) Level
2.3. Mitochondrial DNA (mtDNA) Copy Number Quantification
2.4. Determination of Mitochondrial DNA Damage
3. Discussion
4. Materials and Methods
4.1. Reagents
4.2. Cell Culture
4.3. Assessment of Mitochondrial Membrane Potential (MMP)
4.4. Assessment of Intracellular ROS Level
4.5. Isolation of Total Genomic DNA from Cell Lines
4.6. Mitochondrial DNA Copy Number Quantification
4.7. Determination of Mitochondrial DNA Damage
4.8. Data Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Target Gene Symbol | Forward Primer Sequences (5′→3′) | Reverse Primer Sequence (5′→3′) | Amplicon Length (bp) |
---|---|---|---|
ND1 (mitochondrially encoded NADH: ubiquinone oxidoreductase core subunit 1) | Long fragment: ATGGCCAACCTCCTACTCCT | Long fragment: GATGAGTGTGCCTGCAAAGA | 1214 |
Small fragment: CCTAAAACCCGCCACATCTA | Small fragment: GCCTAGGTTGAGGTTGACCA | 124 | |
ND5 (mitochondrially encoded NADH: ubiquinone oxidoreductase core subunit 5) | Long fragment: TCCAACTCATGAGACCCACA | Long fragment: AGGTGATGATGGAGGTGGAG | 1156 |
Small fragment: AGGCGCTATCACCACTCTGT | Small fragment: TTGGTTGATGCCGATTGTAA | 124 |
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Janik-Karpinska, E.; Ceremuga, M.; Niemcewicz, M.; Synowiec, E.; Sliwiński, T.; Bijak, M. Mitochondrial Damage Induced by T-2 Mycotoxin on Human Skin—Fibroblast Hs68 Cell Line. Molecules 2023, 28, 2408. https://doi.org/10.3390/molecules28052408
Janik-Karpinska E, Ceremuga M, Niemcewicz M, Synowiec E, Sliwiński T, Bijak M. Mitochondrial Damage Induced by T-2 Mycotoxin on Human Skin—Fibroblast Hs68 Cell Line. Molecules. 2023; 28(5):2408. https://doi.org/10.3390/molecules28052408
Chicago/Turabian StyleJanik-Karpinska, Edyta, Michal Ceremuga, Marcin Niemcewicz, Ewelina Synowiec, Tomasz Sliwiński, and Michal Bijak. 2023. "Mitochondrial Damage Induced by T-2 Mycotoxin on Human Skin—Fibroblast Hs68 Cell Line" Molecules 28, no. 5: 2408. https://doi.org/10.3390/molecules28052408