(a)
(b)
(c)
Figure 5: Effects of vitamin D treatment on ROS generation and caspase-3 protein expression 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. (a) The effects of vitamin D treatment on intracellular ROS levels in HUVECs were observed by fluorescence microscope. (b) The effects of vitamin D treatment on intracellular ROS levels in HUVECs were measured by flow cytometry. 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). (c) Effects of vitamin D treatment on caspase-3 protein expression in high glucose-cultured HUVECs. Caspase-3 protein expression levels were expressed as the ratio of caspase-3 over β-actin. HG: high glucose (33 mM); results are presented as mean ± SEM (); versus control; ; versus HG (33 mM).