title: Detection of Modified Forms of Cytosine Using Sensitive Immunohistochemistry.
abstract: Methylation of cytosine bases (5-methylcytosine, 5mC) occurring in vertebrate genomes is usually associated with transcriptional silencing. 5-hydroxylmethylcytosine (5hmC), 5-formylcytosine (5fC), and 5-carboxylcytosine (5caC) are the recently discovered modified cytosine bases produced by enzymatic oxidation of 5mC, whose biological functions remain relatively obscure. A number of approaches ranging from biochemical to antibody based techniques have been employed to study the genomic distribution and global content of these modifications in various biological systems. Although some of these approaches can be useful for quantitative assessment of these modified forms of 5mC, most of these methods do not provide any spatial information regarding the distribution of these DNA modifications in different cell types, required for correct understanding of their functional roles. Here we present a highly sensitive method for immunochemical detection of the modified forms of cytosine. This method permits co-detection of these epigenetic marks with protein lineage markers and can be employed to study their nuclear localization, thus, contributing to deciphering their potential biological roles in different experimental contexts.
authors: Laboratoire de Neurophysiologie (CP601), ULB Neuroscience Institute (UNI), Université Libre de Bruxelles.
authors: Abdulkadir
authors: A
authors: Abakir
authors: Medical Molecular Sciences, Centre for Biomolecular Sciences, University of Nottingham.
authors: Lee
authors: L
authors: Wheldon
authors: School of Life Sciences, University of Nottingham.
authors: Andrew D
authors: AD
authors: Johnson
authors: Laboratoire de Neurophysiologie (CP601), ULB Neuroscience Institute (UNI), Université Libre de Bruxelles.
authors: Patrick
authors: P
authors: Laurent
authors: Division of Cancer and Stem Cells, Centre for Biomolecular Sciences, School of Medicine, University of Nottingham; Alexey.Ruzov@nottingham.ac.uk.
authors: Alexey
authors: A
authors: Ruzov
mesh_terms: D044503:5-Methylcytosine; D000906:Antibodies; D003596:Cytosine / Q000737:chemistry*; D019175:DNA Methylation; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D010084:Oxidation-Reduction
chemical_list: D000906:Antibodies; D044503:5-Methylcytosine; D003596:Cytosine