(a)
(b)
Figure 4: Effects of vitamin D treatment on p66Shc phosphorylation and mitochondrial translocation in high glucose-cultured HUVECs. HUVECs were inoculated in a 6-well plate, 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 or cytoplasmic/mitochondrial protein were extracted for immunoblotting analysis. (a) Effects of vitamin D treatment on p66Shc phosphorylation in high glucose-cultured HUVECs. The p66Shc phosphorylation of HUVECs was expressed as the ratio of p-p66Shc over p66Shc. HG: high glucose (33 mM); results are presented as mean ± SEM (); versus control; versus HG (33 mM). (b) Effects of vitamin D treatment on p66Shc mitochondrial translocation in high glucose-cultured HUVECs. The mitochondrial and cytoplasmic p66Shc levels were expressed as Mito-p66Shc/COX4 and Cyto-p66Shc/β-actin, respectively. HG: high glucose; Mito-p66Shc: mitochondria p66Shc; Cyto-p66Shc: cytoplasm p66Shc; COX4: cytochrome c oxidase subunit 4 (mitochondrial internal protein); results are presented as mean ± SEM (); versus control; versus HG (33 mM).