Figure 8
(A,C) Representative western blot of eNOS dimer/monomer distribution in unboiled samples (top blot) and GAPDH or α-tubulin (bottom blot; loading control) in human umbilical vein endothelial cells (HUVECs) treated with methyl-β-cyclodextrin (10 mM dextrin for 60 min) or not (Control). (B,D) Basal changes in eNOS dimer and monomer induced by dextrin, in the presence of tetrahydrobiopterin (BH4, 100 μM), L-arginine (L-arg, 1 mM), or BH4 plus L-arg or not. We processed images of blots changing brightness and contrast and we applied equally over the entire image and controls. (E,F) NO determination with the DAF-2DA probe in HUVECs treated with dextrin or not (Control) in the presence of BH4, L-arg, L-NAME (1 mM), or BH4 plus L-arg. (G) Reactive oxygen species (ROS) detected with the DHE probe in the presence of BH4, L-arg, BH4 plus L-arg, L-NAME, or A23187 (5 μM) in HUVECs treated with dextrin or not. Bars are reported as the mean ± SEM of independent experiments (n = 4–5). Values of blank were not use to perform statistics. *p < 0.05 statistical differences between Dextrin versus control group. #p < 0.05 between dextrin (in the presence of BH4, or L-arg, or BH4 plus L-arg) and only BH4 versus Dextrin group. &p < 0.05 between L-NAME, dextrin plus L-NAME or A23187 versus Control group.
