PIM1 Promotes Survival of Cardiomyocytes by Upregulating c-Kit Protein Expression
Abstract
:1. Introduction
2. Materials and Methods
2.1. Animal Experiments
2.2. Isolation and Culture of Adult Mouse Cardiomyocytes
2.3. Viral Infections
2.4. Pharmacological Treatments
2.5. Immunoblot
2.6. Quantitative RT-PCR
- c-Kit Forward: ATTGTGCTGGATGGATGGAT
- c-Kit Reverse: GATCTGCTCTGCGTCCTGTT
- Co-Immunoprecipitation
2.7. Proximity Ligation Assay
2.8. Cell Death Assay
2.9. Statistical Analyses
3. Results
3.1. PIM1 Upregulates c-Kit Protein Expression
3.2. PIM1 Interacts with c-Kit
3.3. Cardioprotective Signaling Downstream of c-Kit is Elevated in PIM1 Cardiomyocytes
3.4. Elevated c-Kit Expression Enhances Resistance Against Oxidative Stress in Cardiomyocytes
3.5. PIM1 Confers Cardioprotection Independent of c-kit Activity
4. Discussion
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Antibody | Catalog Number | Dilution | Application |
---|---|---|---|
PIM1 | ThermoFisher 39-4600 | 1:500 | IB, Co-IP |
PIM1 | ThermoFisher 710504 | 1:100 | PLA |
c-Kit | R&D AF1356 | 1:200 1:100 | IB, PLA |
GAPDH (Glyceraldehyde 3-phosphate) dehydrogenase | Millipore Sigma MAB374 | 1:5000 | IB |
SRC | Abcam Ab47405 | 1:500 | IB |
pERK1/2 Phosphorylated ERK1/2 | CST 9101 | 1:500 | IB |
ERK1/2 | CST 9102 | 1:500 | IB |
pAkt (Ser473) Phosphorylated Akt at serine 473 | CST 9271 | 1:500 | IB |
Akt | CST 9272 | 1:500 | IB |
GFP (Green Fluorescent Protein) | Rockland 600-101-215 | 1:1000 | IB |
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Ebeid, D.E.; Firouzi, F.; Esquer, C.Y.; Navarrete, J.M.; Wang, B.J.; Gude, N.A.; Sussman, M.A. PIM1 Promotes Survival of Cardiomyocytes by Upregulating c-Kit Protein Expression. Cells 2020, 9, 2001. https://doi.org/10.3390/cells9092001
Ebeid DE, Firouzi F, Esquer CY, Navarrete JM, Wang BJ, Gude NA, Sussman MA. PIM1 Promotes Survival of Cardiomyocytes by Upregulating c-Kit Protein Expression. Cells. 2020; 9(9):2001. https://doi.org/10.3390/cells9092001
Chicago/Turabian StyleEbeid, David E., Fareheh Firouzi, Carolina Y. Esquer, Julian M. Navarrete, Bingyan J. Wang, Natalie A. Gude, and Mark A. Sussman. 2020. "PIM1 Promotes Survival of Cardiomyocytes by Upregulating c-Kit Protein Expression" Cells 9, no. 9: 2001. https://doi.org/10.3390/cells9092001