Tebuconazole Induces ER-Stress-Mediated Cell Death in Bovine Mammary Epithelial Cell Lines
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture and Treatment
2.2. Cytotoxicity Tests
2.3. Flow Cytometry
2.4. Immunostaining
2.5. RNA Isolation and Quantitative PCR
2.6. Western Blotting
2.7. Statistical Analysis
3. Results
3.1. TEB Induced MAC-T Cell Apoptosis and Toxicity
3.2. Anti-Proliferation Effect of TEB on MAC-T Cell Culture
3.3. TEB Induces Mitochondrial Dysfunction in MAC-T Cells
3.4. TEB Induced Apoptosis via ER Stress in MAC-T Cells
3.5. TEB Regulated the Expression of Milk-Protein-Synthesis-Related Genes and Inflammatory Genes in MAC-T Cells
4. Discussion
5. Conclusions
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
References
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Gene | Forward Primer | Reverse Primer |
---|---|---|
GAPDH | 5′-GGGTCATCATCTCTGCACCT-3′ | 5′-GGTCATAAGTCCCTCCACGA-3′ |
CHOP | 5′-GCAACGCATGAAGGAGAAAG-3′ | 5′-AACCATCCGGTCAATCAGAG-3′ |
GRP78 | 5′-TGGCTGGAAAGTCACCAAG-3′ | 5′-GTCTGCTGCTTCCTCCTCAC-3′ |
ATF4 | 5′-GCTGTGGATTGGTTGGTCTC-3′ | 5′-AGCTCATCTGGCAT-3′ |
LGB | 5′-CTTGTGCTGGACACCGACTA-3′ | 5′-TTGAGGGCTTTGTCGAATTT-3′ |
LALA | 5′-AAAGACGACCAGAACCCTCA-3′ | 5′-GCTTTATGGGCCAACCAGTA-3′ |
CSN1S1 | 5′-CACTGAGTCAAAGGGAATTAAAG-3′ | 5′-TGATGGCACTTACAGGAGA-3′ |
CSN1S2 | 5′-CCTAACAGCCTCCCACAAAA-3′ | 5′-AGACTGGAGCAGAGGCAGAG-3′ |
CSNK | 5′-CCAGGAGCAAAACCAAGAAC-3′ | 5′-TGCAACTGGTTTCTGTTGGT-3′ |
TGFB3 | 5′-TCTGGGGCGACTTAAGAAGA-3′ | 5′-ATTGCGGAAGCAGTAATTGG-3′ |
CEBPD | 5′-ATCGACTTCAGCGCCTACAT-3′ | 5′-TGTGGTTGCTGTTGAAGAGG-3′ |
IL6 | 5′-AAGCAGCAAGGAGACACTGG-3′ | 5′-GCCTGATTGAACCCAGATTG-3′ |
Antibody | Manufacturer | Catalog Number | Dilution (Usage) |
---|---|---|---|
Cleaved-caspase3 | Cell signaling(Danvers, MA, USA) | #9664S | 1:2000 (WB) |
Cleaved-capase8 | Cell signaling | #8592S | 1:2000 (WB) |
BAX | Santa Cruz(Dallas, Texas, USA) | Sc-7480 | 1:2000 (WB) |
BIP/GRP78 | Cell signaling | #3177 | 1:2000 (WB) |
ERO-La | Cell signaling | #377009 | 1:2000 (WB) |
PDI | Cell signaling | #3501 | 1:2000 (WB) |
β-actin | Santa Cruz | Ab16667 | 1:2000 (WB) |
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Lee, W.-Y.; Lee, R.; Park, H.-J. Tebuconazole Induces ER-Stress-Mediated Cell Death in Bovine Mammary Epithelial Cell Lines. Toxics 2023, 11, 397. https://doi.org/10.3390/toxics11040397
Lee W-Y, Lee R, Park H-J. Tebuconazole Induces ER-Stress-Mediated Cell Death in Bovine Mammary Epithelial Cell Lines. Toxics. 2023; 11(4):397. https://doi.org/10.3390/toxics11040397
Chicago/Turabian StyleLee, Won-Young, Ran Lee, and Hyun-Jung Park. 2023. "Tebuconazole Induces ER-Stress-Mediated Cell Death in Bovine Mammary Epithelial Cell Lines" Toxics 11, no. 4: 397. https://doi.org/10.3390/toxics11040397