Effect Smad2C phosphorylation on endothelial cells treated with 15 mM D-glucose for up to 4 h. A. Western blot of whole cell lysates from BAECs treated with 15 mM D-glucose for up to 240 min. Cells treated with TGF-β (2 ng/ml) (5 min) were used as positive control. Cell lysate were resolved on SDS-PAGE and transferred onto a PVDF membrane. The membrane was probed with anti-phospho-Smad2(Ser465/467) antibody. Also probed with anti-Smad2 and anti-GAPDH. Chemiluminescence was used to detect the proteins of interst. B. Histogram is a denstitometric quantitation of. Results determined by one-way ANOVA **p < 0.01 treatments versus basal n = 3 experiments.