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. Author manuscript; available in PMC: 2010 Dec 1.
Published in final edited form as: Oncogene. 2010 Apr 26;29(24):3532–3544. doi: 10.1038/onc.2010.124

Figure 5. K1 protein expression is dependent on Hsp90 activity and the endogenous levels of Hsp90β and ER-associated Hsp40.

Figure 5

A. Stable 293-Vec cells expressing empty control vector (pcDNA3) or 293-K1 cells expressing FLAG-K1 were treated with 1 µM of the Hsp90 inhibitor, geldanamycin (GA), or DMSO (vehicle), for 6 h in the absence of serum. Cell lysates were harvested and resolved by SDS-PAGE and immunoblotting. Western blots were probed with the indicated antibodies. Arrows indicate monomeric and multimeric forms of the K1 protein. B. K1 does not affect the endogenous levels of Hsp90β and Hsp40. Stable 293-K1 or 293-Vec cell lysates were subjected to SDS-PAGE and Western blot analysis using the indicated antibodies. Each panel is representative of at least three independent experiments. C. 293-K1 cells were subjected to siRNA targeting luciferase (siLUC; control), or Hsp90β (siHsp90β-1 or siHsp90β-1&2) for 48 h. Cell lysates were harvested and subjected to SDS-PAGE and Western blot analysis. D. 293-K1 cells were subjected to siRNA targeting luciferase (siLUC; control) or a pool of siRNAs targeting ER-associated Hsp40 (siHsp40) for 48 h. Cell lysates were harvested and subjected to SDS-PAGE and Western blot analysis.