Glucolipotoxic Stress-Induced Mig6 Desensitizes EGFR Signaling and Promotes Pancreatic Beta Cell Death
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture, Reagents, and the Use of Adenovirus
2.2. Human and Rodent Islet Experiments
2.3. Apoptosis Assays
2.4. Immunoblot Analysis
2.5. Quantitative RT-PCR Analysis
2.6. Statistical Analysis
3. Results
3.1. GLT and ER Stress Attenuate EGFR Activation in Rodent Beta Cells and Human Islets
3.2. EGFR Feedback Inhibitor Mig6 Is Elevated in GLT-Treated Beta Cells and T2D Human Islets
3.3. Mig6 Inhibits EGFR in Pancreatic Beta Cells and Promotes Death during GLT
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Name | Vendor, Model Number | Dilution |
---|---|---|
Anti-Actin | MP Biomedicals, #691002 | 1:5000 |
Anti-Akt | Cell Signaling, #2920 | 1:1000 |
Anti-caspase 3 | Cell Signaling, #9662 | 1:1000 |
Anti-CHOP | Santa Cruz, #7351 | 1:250 |
Anti-EGFR | Sigma-Aldrich, #E3138 | 1:1000 |
Anti-eIF2α | Cell Signaling, #5324 | 1:1000 |
Anti-ERK1/2 | Cell Signaling, #4696 | 1:1000 |
Anti-γ-tubulin | Sigma-Aldrich, #T6557 | 1:5000 |
Anti-GAPDH | Abcam, #Ab9483 | 1:5000 |
Anti-Mig6 | Santa Cruz, #D-1 | 1:250 |
Anti-phospho-Akt (Thr308) | Cell Signaling, #4056 | 1:1000 |
Anti-phospho-EGFR (Tyr1068) | Cell Signaling, #3777 | 1:1250 |
Anti-phospho-eIF2α (Ser51) | Cell Signaling, #3398 | 1:1000 |
Anti-phospho-ERK1/2 (Thr202/Tyr204) | Cell Signaling, #4370 | 1:2000 |
IRDye 800 or 700 fluorophore-conjugated antibodies | LI-COR | 1:10,000 |
Name | Vendor | Assay ID |
---|---|---|
Rat Mig6 | ThermoFisher Scientific | Rn01520744_g1 |
Human errfi1 (Mig6) | ThermoFisher Scientific | Hs00219060_m1 |
Rat Socs4 | ThermoFisher Scientific | Rn01414734_m1 |
Rat Socs5 | ThermoFisher Scientific | Rn01769079_m1 |
Rat Frs3 | ThermoFisher Scientific | Rn01512038_m1 |
Rat Lrig1 | ThermoFisher Scientific | Rn01421201_m1 |
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Chen, Y.-C.; Lutkewitte, A.J.; Basavarajappa, H.D.; Fueger, P.T. Glucolipotoxic Stress-Induced Mig6 Desensitizes EGFR Signaling and Promotes Pancreatic Beta Cell Death. Metabolites 2023, 13, 627. https://doi.org/10.3390/metabo13050627
Chen Y-C, Lutkewitte AJ, Basavarajappa HD, Fueger PT. Glucolipotoxic Stress-Induced Mig6 Desensitizes EGFR Signaling and Promotes Pancreatic Beta Cell Death. Metabolites. 2023; 13(5):627. https://doi.org/10.3390/metabo13050627
Chicago/Turabian StyleChen, Yi-Chun, Andrew J. Lutkewitte, Halesha D. Basavarajappa, and Patrick T. Fueger. 2023. "Glucolipotoxic Stress-Induced Mig6 Desensitizes EGFR Signaling and Promotes Pancreatic Beta Cell Death" Metabolites 13, no. 5: 627. https://doi.org/10.3390/metabo13050627