Fig.3.

WA and its analogues direct bind to Hsp90 and induce Hsp90 aggregation. A. WA, WE, HWE, AzWA and WP compete with WA-biotin binding to Hsp90. One mg cell lysate was preincubated with 100μM WA or its analogues for 1 h, respectively before subject to WA-biotin pull down assay. The WA-biotin pull down protein were subjected to Western blot analysis with specific antibodies to Hsp90. B. WA, WE, HWE or AzWA induces Hsp90 aggregation. Panc-1 cells were treated with 10 μM WA or its analogues for 24 h, respectively. Equal amounts of protein (50μg/lane) were subjected to non-reducing gel electrophoresis and then analyzed by Western blot with specific antibodies to Hsp90 and Actin, Actin was served as internal standard. Results are representative of three independent experiments.