Prevention of High Glucose-Mediated EMT by Inhibition of Hsp70 Chaperone
Abstract
:1. Introduction
2. Results
2.1. Expression of Hsp70 Links to the Capacity of Hyperglycemia-Induced EMT
2.2. Suppressing Hsp70 Reduces the Ability of Colon Cancer Cells to Pass EMT under Hyperglycemia Conditions
2.3. Inhibition of Chaperonic Function Leads to Downregulation of EMT in Colon Cancer Cells
3. Materials and Methods
3.1. Cells and Treatment
3.2. Wound Healing Assay
3.3. Chemicals
3.4. Western Blotting
3.5. RNA Isolation and Quantitative Real-Time PCR
3.6. Immunocytochemistry
3.7. Chap-ELISA
3.8. Statistics
4. Discussion
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Antibodies to: | Manufacturer | Catalogue Number |
---|---|---|
Vimentin | Abcam | ab137321 |
E-cadherin | Abcam | ab40772 |
Hsp70 | Homemade | clone 2E4 |
GAPDH | Thermo Fisher | AM4300 |
ß-tubulin | Thermo Fisher | MA1-80017 |
HSF1 | Cell Signaling | #4356 |
GAM | Abcam | ab6789 |
GARabbit | Abcam | ab6721 |
GARat | Sigma-Aldrich (Merck) | AP136P |
Goat pAb to Ms IgG Alexa647 | Abcam | ab150115 |
β-actin | Forward | CCATCATGAAGTGTGACGTGG |
Reverse | GTCCGCCTAGAAGCATTTGCG | |
Snail | Forward | CATCCTTCTCACTGCCATG |
Reverse | GTCTTCATCAAAGTCCTGTGG | |
E-cadherin | Forward | AGGCCAAGCAGCAGTACATT |
Reverse | ATTCACATCCAGCACATCCA | |
Slug | Forward | ATGAGGAATCTGGCTGCTGT |
Reverse | CAGGAGAAAATGCCTTTGGA | |
Twist | Forward | TGCATGCATTCTCAAGAGGT |
Reverse | CTATGGTTTTGCAGGCCAGT |
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Nikotina, A.D.; Vladimirova, S.A.; Komarova, E.Y.; Alexeev, D.; Efremov, S.; Leonova, E.; Pavlov, R.; Kartsev, V.G.; Polonik, S.G.; Margulis, B.A.; et al. Prevention of High Glucose-Mediated EMT by Inhibition of Hsp70 Chaperone. Int. J. Mol. Sci. 2021, 22, 6902. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22136902
Nikotina AD, Vladimirova SA, Komarova EY, Alexeev D, Efremov S, Leonova E, Pavlov R, Kartsev VG, Polonik SG, Margulis BA, et al. Prevention of High Glucose-Mediated EMT by Inhibition of Hsp70 Chaperone. International Journal of Molecular Sciences. 2021; 22(13):6902. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22136902
Chicago/Turabian StyleNikotina, Alina D., Snezhana A. Vladimirova, Elena Y. Komarova, Dmitry Alexeev, Sergey Efremov, Elizaveta Leonova, Rostislav Pavlov, Viktor G. Kartsev, Sergey G. Polonik, Boris A. Margulis, and et al. 2021. "Prevention of High Glucose-Mediated EMT by Inhibition of Hsp70 Chaperone" International Journal of Molecular Sciences 22, no. 13: 6902. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22136902