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Whether nucleoskeletons seen after extracting cells are preparative artefacts is controversial. Using an extraction method that preserves vital nuclear functions, we have visualized part of a nucleoskeleton by electron microscopy of thick resinless sections. Cells encapsulated in agarose microbeads are lysed using Triton in a physiological buffer; the agarose coat prevents aggregation and protects fragile cell contents. These extracted cells are accessible to small molecules and transcribe and replicate at rates close to those in vivo. After electroeluting most chromatin after treatment with HaeIII, a skeleton is uncovered which ramifies throughout the nucleus. Individual filaments are approximately 10 nm wide with an axial repeat of 23 nm, characteristic of intermediate filaments.

Type

Journal article

Journal

EMBO J

Publication Date

01/12/1988

Volume

7

Pages

3667 - 3677

Keywords

Cell Nucleus, Chromatin, Deoxyribonucleases, Electrophoresis, HeLa Cells, Microscopy, Electron, Nuclear Proteins, Ribonucleases