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. Author manuscript; available in PMC: 2008 Jan 15.
Published in final edited form as: Free Radic Biol Med. 2006 Oct 12;42(2):191–201. doi: 10.1016/j.freeradbiomed.2006.10.036

Figure 4.

Figure 4

Effect of arsenite on Hsp90 expression, ubiquitination, and association with co-chaperone and client proteins. (A) Effect of arsenite on the steady-state levels of Hsp90. Hsp90 was immunoprecipitated from total cell lysates with anti-Hsp90 antibody and immunoblotted with the same antibody. (B) Effect of arsenite on Hsp90 ubiquitination in GCS-2 cells. Hsp90 was immunoprecipitated with anti-Hsp90, separated by SDS-PAGE and analyzed by immunoblot with anti-ubiquitin antibody. (C) Effect of arsenite on Hsp90 association with its cochaperone p50cdc37. Cell lysates were immunoprecipitated with anti-Hsp90 antibody and blotted with anti-p50cdc37 antibody. (D) Effect of arsenite on steady-state protein levels of Hsp70. Cell lysates were immunoprecipitated with anti-Hsp70 antibody and immunoblotted with the same antibody. This also serves as a loading control. (E) Effect of arsenite on Plk-1 expression in GCS-2 cells. Total cell lysates were immunoblotted with anti-Plk-1 antibody. (F) Effect of arsenite on cyclin D-associated cdk-4 expression in GCS-2 cells. Plk-1 membrane was stripped and immunoblotted using anti-cdk-4 antibody. (G) Cdk-4 membrane was stripped and reprobed with actin-specific antibody to control for protein loading.