Trichostatin A-Assisted Epigenomic Modulation Affects the Expression Profiles of Not Only Recombinant Human α1,2-Fucosyltransferase and α-Galactosidase A Enzymes But Also Galα1→3Gal Epitopes in Porcine Bi-Transgenic Adult Cutaneous Fibroblast Cells
Abstract
:1. Introduction
2. Results
2.1. Western Blot Analysis of the Relative Expression of Recombinant Human α1,2-Fucosyltransferase (rhα1,2-FT) and α-Galactosidase A (rhα-Gal A) Proteins
2.2. Immunofluorescence Localization of Recombinant Human α1,2-Fucosyltransferase (rhα1,2-FT) and α-Galactosidase A (rhα-Gal A) in Trichostatin A-Treated and Untreated ACFCs
2.3. Lectin Blotting Analysis of Galα1→3Gal Epitope Expression at the Protein Level in the In Vitro-Cultured Porcine Bi-Transgenic and Non-Transgenic ACFCs Treated and Not Treated with Trichostatin A
2.4. Identification of the Expression Profiles of Galα1→3 Gal Epitope by Fluorescently Labelled Lectin GS-IB4
3. Discussion
4. Materials and Methods
4.1. In Vitro Culture and Trichostatin A-Mediated Epigenomic Modulation of Fibroblast Cells
4.2. Total Protein Extraction and Western/Lectin Blot Analyses
4.3. Immunofluorescence Staining
4.4. Lectin Fluorescence
4.5. Confocal Microscope Analyses
4.6. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
Abbreviations
ACFCs | Adult cutaneous fibroblast cells |
DNMTi | Inhibitors of DNA methyltransferases |
HAR | Hyperacute rejection |
HDACi | Inhibitors of histone deacetylases |
HRP | Horseradish peroxidase |
iPSCs | Induced pluripotent stem cells |
MSCs | Mesenchymal stem cells |
rhα1,2-FT | Recombinant human α1,2-fucosyltransferase |
rhα-Gal A | Recombinant human α-galactosidase A |
SCNT | Somatic cell nuclear transfer |
TSA | Trichostatin A |
α1,3GT | α1,3-galactosyltransferase |
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Wiater, J.; Samiec, M.; Skrzyszowska, M.; Lipiński, D. Trichostatin A-Assisted Epigenomic Modulation Affects the Expression Profiles of Not Only Recombinant Human α1,2-Fucosyltransferase and α-Galactosidase A Enzymes But Also Galα1→3Gal Epitopes in Porcine Bi-Transgenic Adult Cutaneous Fibroblast Cells. Int. J. Mol. Sci. 2021, 22, 1386. https://doi.org/10.3390/ijms22031386
Wiater J, Samiec M, Skrzyszowska M, Lipiński D. Trichostatin A-Assisted Epigenomic Modulation Affects the Expression Profiles of Not Only Recombinant Human α1,2-Fucosyltransferase and α-Galactosidase A Enzymes But Also Galα1→3Gal Epitopes in Porcine Bi-Transgenic Adult Cutaneous Fibroblast Cells. International Journal of Molecular Sciences. 2021; 22(3):1386. https://doi.org/10.3390/ijms22031386
Chicago/Turabian StyleWiater, Jerzy, Marcin Samiec, Maria Skrzyszowska, and Daniel Lipiński. 2021. "Trichostatin A-Assisted Epigenomic Modulation Affects the Expression Profiles of Not Only Recombinant Human α1,2-Fucosyltransferase and α-Galactosidase A Enzymes But Also Galα1→3Gal Epitopes in Porcine Bi-Transgenic Adult Cutaneous Fibroblast Cells" International Journal of Molecular Sciences 22, no. 3: 1386. https://doi.org/10.3390/ijms22031386