Premium
Basic fibroblast growth factor accumulates in the nuclei of various bFGF‐producing cell types
Author(s) -
Tessler Shoshanna,
Neufeld Gera
Publication year - 1990
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1041450216
Subject(s) - basic fibroblast growth factor , transfection , microbiology and biotechnology , biology , polyclonal antibodies , baby hamster kidney cell , recombinant dna , intracellular , antibody , blot , cell culture , cell , growth factor , biochemistry , immunology , gene , receptor , genetics
The intracellular localization of basic fibroblast growth factor (bFGF) was studied in BHK‐21 cells transfected with an expression vector containing the complementary DNA (cDNA) of the human bFGF gene (pbFGF). The intracellular location of bFGF was determined using indirect immunofluorescence. The antibodies used were polyclonal antibodies directed against either recombinant human bFGF or recombinant Xenopus : bFGF. The nuclei of transfected cells that produce bFGF, but not the nuclei ofuntransfeted cells, were labeled strongly by the antibodies. The nuclear staining was totally abolished when anti‐bFGF antibodies preadsorbed with bFGF were used. Several types of endothelial cells known to produce bFGF were also stained in their nuclei by the antibodies. Nuclear extracts prepared from transfected cells were found to contain bFGF as determined using heparin‐sepharose affinity chromatography, followed by Western blot analysis of fractions, which stimulated the proliferation BHK‐21 cells. The mitogenic cactivity associated with the nuclei was not destroyed when isolated cell nuclei were digested by trypsin. It is therefore likely that the nucleus associated bFGF is intranuclear, these findings suggest that some biologicaL activities of bFGF may be mediated by nuclear bFGF binding proteins or by the direct binding of bFGF‐ to DNA.