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. 2012 Dec 21;14(1):255–272. doi: 10.3390/ijms14010255

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© 2013 by the authors; licensee Molecular Diversity Preservation International, Basel, Switzerland.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).

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CrAT gene knockdown disrupts eNOS/Hsp90 interactions in pulmonary arterial endothelial cells. PAEC were transiently transfected with a scrambled siRNA or a specific CrAT siRNA for 48 h. Western blot analysis revealed that reducing CrAT expression did not alter total protein levels of (A) eNOS or (B) Hsp90 (C) However, immunoprecipitation (IP) analysis using a specific antiserum raised against Hsp90 followed by Western blot (IB) analysis with an anti-eNOS antibody revealed that there was a significant decrease in the association of eNOS with Hsp90 in CrAT siRNA transfected cells. The membrane was then reprobed for Hsp90 to normalize for immunoprecipitation efficiency. Values are mean ± SEM; N = 6. *p < 0.05 vs. scrambled siRNA.