FIG. 3.

HSP90 and HSP70 are eluted from the affinity chromatography with DEN virus recombinant E protein. Thirty micrograms of total protein extract from SK-SY-5Y (lanes 1) and U937 cells (lanes 3), 50 μl of eluted fraction from SK-SY-5Y (lanes 2) and U937 cells (lanes 4) obtained in the presence of recombinant E protein, or 50 μl of eluted fraction from SK-SY-5Y (lanes C) and U937 cells (lanes C') obtained in the absence of recombinant E protein was separated by SDS-10% PAGE, transferred to nitrocellulose membrane, and incubated with monoclonal antibodies against HSP90 (A) or anti-HSP70 (B). Then membranes were incubated with a second antibody goat anti-mouse IgG coupled to peroxidase. Reactions were developed by chemiluminescence. Molecular mass markers are indicated on the left. Arrows indicate migration of HSP90 and HSP70.