Fig. 3.

Identification of vimentin cleavage fragments as HSP90 binding protein. a Subcellular colocalization of vimentin and Hsp90. Cells were doubly stained for vimentin and Hsp90. b Adherent growing HepG2 cells were kept either untreated or were irradiated (500 μM H₂O₂, 24 h). Immediately after oxidative stress, cells were lysed and soluble and insoluble vimentin were assayed by Western blotting using anti-vimentin. Equal amounts (20 μg) of protein were loaded. c HepG2 cells were immunoprecipitated with anti-Hsp90 antibodies. The resulting pellets were subjected to SDS-PAGE and analyzed by immunoblotting with anti-vimentin antibodies. The signal intensity obtained from HepG2 cells without added GA was arbitrarily set to 1.0 (control)