Inhibition of Hsp90 resulted in eNOS monomerization without change phosphorylation status of Serine 1179 and Threonine 497 in eNOS dimmer.

(A) BAECs were treated with 20 µM geldanamycin for 48 hours and the samples were run SDS-PAGE gel at low temperature and blotted with the indicated antibodies to determine the changes in phosphorylation status of eNOS and dimers and monomers. The eNOS dimer and monomer densities were determined using AlphaImager. And The Ser1179 and Thr497 phosphorylation density on eNOS dimer were also determined using AlphaImager. All results are representative of three independent experiments. * and # P<0.05 comparison with control. (B) BAECs were silenced with scramble siRNA control (scRNA) or Hsp90 (siHsp90) as indicated concentration for 3 days, the cell lysates were blotted with eNOS or Hsp90 antibody respectively. The eNOS monomer and dimer density were measured using AlphaImager and the average of three independent experiments was presented (* and #P<0.05 compared to eNOS dimer and monomer control respectively).