Figure 7: Effects of vitamin D treatment on eNOS protein expression and activity and NO generation in high glucose-cultured HUVECs. HUVECs were inoculated in 6-well plates, cultured with 5% FBS at 70%~80% confluence for 24 h, and then coincubated with high glucose (33 mM) and vitamin D (10−6 M) or Juglone (10−7 M) for 72 h. Total proteins were extracted for immunoblotting analysis. The eNOS protein expression levels and activity were expressed as the ratio of eNOS over β-actin and p-eNOS over β-actin, respectively. The NO generation of HUVEC was measured by Griess method. HG: high glucose (33 mM); results are presented as mean ± SEM (); versus control; versus HG (33 mM); versus HG (33 mM) + VitD (10−6 M).