Taxol, a potent inhibitor of human HeLa and mouse fibroblast cell replication, blocked cells in the G2 and M phase of the cell cycle and stabilized cytoplasmic microtubules. The cytoplasmic microtubules of taxol-treated cells were visualized by transmission electron microscopy and indirect immunofluorescence microscopy. More than 90% of the cells treated with 10 micro M taxol for 22 hr at 37 degrees C displayed bundles of microtubules that appeared to radiate from a common site (or sites), in addition to their cytoplasmic microtubules. Untreated cells that were kept in the cold (4 degrees C) for 16 hr lost their microtubules, whereas cells that were pretreated with taxol for 22 hr at 37 degrees C continued to display their microtubules and bundles of microtubules in the cold. Taxol inhibited the migration behavior of fibroblast cells, but these cells did not lose their ability to produce mobile surface projections such as lamellipodia and filopodia.

https://www.pnas.org/content/pnas/77/3/1561.full.pdf

Taxol stabilizes microtubules in mouse fibroblast cells.

A cytoskeletal structure with associated polyribosomes obtained from HeLa cells.

Antibody to prekeratin: Decoration of tonofilament-like arrays in various cells of epithelial character

Publisher Summary This chapter describes the study of the intracellular location of proteins by immunocytochemical procedures. Although the methods given in the chapter are generally applicable, they are documented for tubulin containing structures and antibodies to tubulin. Further examples of the usefulness of these methods in studying cell structure can be found in several recent reviews. Immunofluorescence microscopy is a very sensitive method, theoretically requiring only one antigenic site on the structural protein. It offers several unique advantages for the study of cytoskeletal structures. First, only the protein of choice is visualized by use of its specific antibody. Second, if the protein forms part of a supramolecular structure, the organization of that structure throughout the cell can be demonstrated. Third, numerous cells can be observed simultaneously, and thus the unique features of the organization under different experimental conditions can be observed. Thus, immunofluorescence microscopy can bridge the gap between light microscopy and electron microscopy.

Immunofluorescence and immunocytochemical procedures with affinity purified antibodies: tubulin-containing structures.

The detergent-resistant cytoskeleton of tissue culture cells includes the nucleus and the microfilament bundles

Specific visualization of tubulin-containing structures in tissue culture cells by immunofluorescence. Cytoplasmic microtubules, vinblastine-induced paracrystals, and mitotic figures.

Th. Bibring

Papers

Specific visualization of tubulin-containing structures in tissue culture cells by immunofluorescence. Cytoplasmic microtubules, vinblastine-induced paracrystals, and mitotic figures.