Newly discovered oxidized forms of 5-methylcytosine (oxi-mCs), 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC) may represent distinct DNA modifications with unique functional roles. Here a semi-quantitative workflow for visualization of oxi-mCs' spatial distribution, signal intensity profiling and colocalization is described.
Ramsawhook, A. H., Lewis, L. C., Eleftheriou, M., Abakir, A., Durczak, P., Markus, R., Rajani, S., Hannan, N. R., Coyle, B., Ruzov, A. Immunostaining for DNA Modifications: Computational Analysis of Confocal Images. J. Vis. Exp. (127), e56318, doi:10.3791/56318 (2017).