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Combined Immunofluorescence and DNA FISH on 3D-preserved Interphase Nuclei to Study Changes in 3D Nuclear Organization
Published on: February 3, 2013
Differential location of nucleic acids within interchromatin granule clusters
1Institute of Cell and Tissue Biology, University of Liège, Belgium.
Abstract:
We have examined in great detail the distribution of nucleic acids within interchromatin granule clusters in different cell types by means of various immunocytochemical approaches. Using the in situ polyadenylate nucleotidyl transferase-immunogold technique for RNA detection or anti-RNA antibodies, we decisively demonstrate the presence of appreciable amount of RNA in clusters of interchromatin granules of untreated cells. Neither the in situ terminal deoxynucleotidyl transferase-immunogold technique nor anti-DNA antibodies, nor the in situ nick-translation technique for DNA detection have revealed any DNA in the interchromatin granule clusters. However, dispersed chromatin sensitive to DNase I are found at the borders and in the close vicinity of interchromatin granule clusters. The results indicate that interchromatin granule clusters should not be nuclear structures directly involved in RNA transcription but rather in some other steps of RNA metabolism.
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