| Chapter title |
The Nucleolus
|
|---|---|
| Chapter number | 1 |
| Book title |
The Nucleolus
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-3792-9_1 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-3790-5, 978-1-4939-3792-9
|
| Authors |
van Sluis, Marjolein, van Vuuren, Chelly, McStay, Brian, Marjolein van Sluis, Chelly van Vuuren, Brian McStay |
| Editors |
Attila Németh |
| Abstract |
3D-immunoFISH is a valuable technique to compare the localization of DNA sequences and proteins in cells where three-dimensional structure has been preserved. As nucleoli contain a multitude of protein factors dedicated to ribosome biogenesis and form around specific chromosomal loci, 3D-immunoFISH is a particularly relevant technique for their study. In human cells, nucleoli form around transcriptionally active ribosomal gene (rDNA) arrays termed nucleolar organizer regions (NORs) positioned on the p-arms of each of the acrocentric chromosomes. Here, we provide a protocol for fixing and permeabilizing human cells grown on microscope slides such that nucleolar proteins can be visualized using antibodies and NORs visualized by DNA FISH. Antibodies against UBF recognize transcriptionally active rDNA/NORs and NOP52 antibodies provide a convenient way of visualizing the nucleolar volume. We describe a probe designed to visualize rDNA and introduce a probe comprised of NOR distal sequences, which can be used to identify or count individual NORs. |
X Demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 15 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 4 | 27% |
| Researcher | 3 | 20% |
| Student > Doctoral Student | 2 | 13% |
| Student > Master | 1 | 7% |
| Student > Bachelor | 1 | 7% |
| Other | 0 | 0% |
| Unknown | 4 | 27% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 5 | 33% |
| Agricultural and Biological Sciences | 4 | 27% |
| Medicine and Dentistry | 1 | 7% |
| Engineering | 1 | 7% |
| Unknown | 4 | 27% |