Over-expression of an active S6K1 mutant causes eNOS uncoupling in endothelial cells.

<p>Young HUVEC cells were transduced either with an empty rAd vector as control or with rAd/CMV-HA-S6K1ca (a constitutively active S6K1 mutant). Two days post transduction, cells were subjected to (<b>A</b>) Immunoblotting analysis of expression of eNOS-dimers, -monomers with anti-eNOS antibody, HA-S6K1-ca with anti-HA antibody. Ponseau S staining served as loading control. Quantification of the eNOS-dimer/monomer ratio is shown in the lower panel. n = 8 in each group. (<b>B</b>) DHE staining was performed 1 hour after L-NAME (1 mmol/L) treatment. Quantification of the signals is shown in the corresponding lower panels. n = 8. **p<0.01 and ***p<0.001 between indicated groups. Scale bar = 0.2 mm.