Original Research ARTICLE
Surv Ophthalmol. Matsumoto Y, Johnson DH. Dexamethasone decreases phagocytosis by human trabecular meshwork cells in situ. Invest Ophthalmol Vis Sci. Gene expression profile of human trabecular meshwork cells in response to long-term dexamethasone exposure. Mol Vis. Glucocorticoids with different chemical structures but similar glucocorticoid receptor potency regulate subsets of common and unique genes in human trabecular meshwork cells.
Regulation of DNA Methylation in Human Breast Cancer
BMC Med Genomics. Chronic corticosterone exposure increases expression and decreases deoxyribonucleic acid methylation of Fkbp5 in mice. Becker B, Hahn KA. Topical corticosteroids and heredity in primary open-angle glaucoma.
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Am J Ophthalmol. Twin study on ocular pressure after topical dexamethasone. Frequency distribution of pressure response. Minireview: epigenetics of obesity and diabetes in humans. Ballestar E. Epigenetic alterations in autoimmune rheumatic diseases.
Nat Rev Rheumatol. Epigenomics of idiopathic pulmonary fibrosis: evaluating the first steps. Ho SM. Environmental epigenetics of asthma: an update.
J Allergy Clin Immunol. A statistical framework for Illumina DNA methylation arrays. Review of processing and analysis methods for DNA methylation array data.
DNA methylation protocols. Overview.
Br J Cancer. Active cytosine demethylation triggered by a nuclear receptor involves DNA strand breaks. Tissue differential microarray analysis of dexamethasone induction reveals potential mechanisms of steroid glaucoma. Substrate rigidity modulates cell matrix interactions and protein expression in human trabecular meshwork cells.
A systems-level integrative framework for genome-wide DNA methylation and gene expression data identifies differential gene expression modules under epigenetic control. Wu H, Zhang Y. Reversing DNA methylation: mechanisms, genomics, and biological functions.
FKbinding protein 51 regulates nuclear transport of the glucocorticoid receptor beta and glucocorticoid responsiveness. Biochemical J. Systemic steroid exposure is associated with differential methylation in chronic obstructive pulmonary disease. Four subunits alphabetagammadelta of the epithelial sodium channel ENaC are expressed in the human eye in various locations. J Exp Zool.
Increased expression of serum amyloid A in glaucoma and its effect on intraocular pressure. DNA methylation-associated colonic mucosal immune and defense responses in treatment-naive pediatric ulcerative colitis. The role of integrins in the trabecular meshwork. J Ocular Pharmacol Ther. Comparative genomic and proteomic analysis of cytoskeletal changes in dexamethasone-treated trabecular meshwork cells. Mol Cell Proteomics.
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Alterations in DNA methylation of Fkbp5 as a determinant of blood-brain correlation of glucocorticoid exposure. Supplement 1. View Metrics. Gel loading buffer see table Gel loading buffer must be added to the samples before loading and serves three main purposes:. Molecular-weight markers should always be included on a gel to enable analysis of DNA fragment sizes in the samples.
See table Commonly used DNA markers in agarose gel electrophoresis for commonly used markers. To allow visualization of the DNA samples, agarose gels are stained with an appropriate dye. The most commonly used dye is the intercalating fluorescent dye ethidium bromide, which can be added either before or after the electrophoresis see table Comparison of ethidium bromide staining methods.
Addition of ethidium bromide prior to electrophoresis — add ethidium bromide at a concentration of 0. Mix the agarose—ethidium bromide solution well to avoid localized staining. Addition of ethidium bromide after electrophoresis — soak the gel in a 0. Tip : Rinse the gel with buffer or water before examining it to remove excess ethidium bromide. Tip : Staining buffer can be saved and re-used. Note : Ethidium bromide is a powerful mutagen and is very toxic. Wear gloves and take appropriate safety precautions when handling.
Use of nitrile gloves is recommended as latex gloves may not provide full protection. After use, ethidium bromide solutions should be decontaminated as described in commonly used manuals 1, 6. Ethidium bromide—DNA complexes display increased fluorescence compared to the dye in solution. This means that illumination of a stained gel under UV light — nm allows bands of DNA to be visualized against a background of unbound dye. The gel image can be recorded by taking a Polaroid photograph or using a gel documentation system.
Tip : UV light can damage the eyes and skin.
DNA Methylation Protocols | SpringerLink
Always wear suitable eye and face protection when working with a UV light source. Southern blotting is a widely used technique that allows analysis of specific DNA sequences. DNA is usually first converted into conveniently sized fragments by restriction digestion. The DNA is next run through an agarose gel 6. Southern blotting named after its inventor, E. Southern refers to the transfer of the DNA to a nylon or nitrocellulose membrane by capillary transfer.