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. Author manuscript; available in PMC: 2010 Sep 8.
Published in final edited form as: Biochemistry. 2009 Sep 8;48(35):8483–8490. doi: 10.1021/bi901058g

FIGURE 2.

FIGURE 2

Hsp90 inhibits basal and Hsp70-stimulated nNOS ubiquitination. A, Hsp90 inhibition of ubiquitination. nNOS was preincubated with Hsp70, Hsp40 and Hsp90 as indicated and then incubated with the ubiquitinating enzyme mixture. The samples were Western blotted by probing with anti-nNOS. Arrow points to the unubiquitinated/monoubiquitinated nNOS band. The relative densities in the bar graph are presented as a % of the Hsp70-stimulated value in lane 3. B, Hsp90 inhibits ubiquitination in the absence and presence of calmodulin. nNOS was preincubated with Hsp70/Hsp40, Hsp90 and 30 μM calmodulin as indicated and then incubated with the ubiquitination mixture, followed by immunoblotting as above. The relative densities in the bar graph are presented as a % of the highest condition in lane 6.