Human Microfibrillar-Associated Protein 4 (MFAP4) Gene Promoter: A TATA-Less Promoter That Is Regulated by Retinol and Coenzyme Q10 in Human Fibroblast Cells
Abstract
:1. Introduction
2. Results
2.1. Prediction and Plasmid Construction of Human MFAP4 Promoter
2.2. Expression of pEGFP-pMFAP4 Plasmids in Human Fibroblast Cells
2.3. Expression of pEGFP-pMFAP4 Plasmids in Mouse Fibroblasts and in Human Melanoma and Mouse Melanoma Cells
2.4. The Alignment of the Human MFAP4 Promoter Region with Those of Other Species
2.5. Effect of Retinol and coQ10 on the Expression of Human MFAP4 Promoter in Detroit 551 Cells
3. Discussion
4. Materials and Methods
4.1. Materials
4.2. Plasmid Construction and Polymerase Chain Reaction (PCR) Experiments
4.3. Cell Lines and Culture Conditions
4.4. Cell Transfection
4.5. Fluorescent Protein Intensity Measurement
4.6. The Alignment of the Human MFAP4 Promoter Region with Those of Other Species
4.7. Cell Viability Assay (MTT Assay)
4.8. Compound Treatments for Promoter Activity Analysis
4.9. SEAP Activity Assay
4.10. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Primer name | Sequence |
---|---|
hMFAP4-P-436-XhoI-F | 5′-gCTCGAGCCCAGAAGCCAGGCATATGA-3′ |
hMFAP4-P-936-XhoI-F | 5′-gCTCGAGGAGGCAATAGGGAGCCATGA-3′ |
hMFAP4-P-1936-XhoI-F | 5′-gCTCGAGTTTACAGAGGAGGAAACCAAGG-3′ |
hMFAP4-P+64-EcoRI-R | 5′-gGAATTCGCTGTCAGTTCTGCTCAGAGT-3′ |
Cell Lines | Species | Features (Tissue/Type) | Elastin Production/hMFAP4 Promoter Activity |
---|---|---|---|
Detroit 551 (BCRC 60118) | Human | Skin/normal/fibroblast | Y/Y |
MeWo (BCRC 60540) | Human | Skin/melanoma | N/N |
NIH/3T3 (BCRC 60008) | Mouse | Embryo/fibroblast | Y/N |
B16-F10 (BCRC 60031) | Mouse | Skin/melanoma | N/N |
MRC-5 (BCRC 60023) | Human | Lung/normal/fibroblast | Y/Y |
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Lin, Y.-J.; Chen, A.-N.; Yin, X.J.; Li, C.; Lin, C.-C. Human Microfibrillar-Associated Protein 4 (MFAP4) Gene Promoter: A TATA-Less Promoter That Is Regulated by Retinol and Coenzyme Q10 in Human Fibroblast Cells. Int. J. Mol. Sci. 2020, 21, 8392. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21218392
Lin Y-J, Chen A-N, Yin XJ, Li C, Lin C-C. Human Microfibrillar-Associated Protein 4 (MFAP4) Gene Promoter: A TATA-Less Promoter That Is Regulated by Retinol and Coenzyme Q10 in Human Fibroblast Cells. International Journal of Molecular Sciences. 2020; 21(21):8392. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21218392
Chicago/Turabian StyleLin, Ying-Ju, An-Ni Chen, Xi Jiang Yin, Chunxiang Li, and Chih-Chien Lin. 2020. "Human Microfibrillar-Associated Protein 4 (MFAP4) Gene Promoter: A TATA-Less Promoter That Is Regulated by Retinol and Coenzyme Q10 in Human Fibroblast Cells" International Journal of Molecular Sciences 21, no. 21: 8392. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21218392