Effect of GKA on D-glucose mediated Smad2C phosphorylation in endothelial cells. A. Western blot of whole cell lysates from BAEC treated with D-glucose 15 mM in the presence and absence of GKA (RO28-1675) (0.1-10 μM). TGF-β was used as positive control. Cell lysates were resolved on SDS-PAGE and transferred onto a PVDF membrane. This membrane was probed with anti-phospho-Smad2(Ser465/467). Also probed for anti-Smad2 and anti-GAPDH. Chemiluminescence was used to detect proteins of interest. B. Histogram is a densitometric quantitation. Results ##p < 0.01 = compared to untreated basal level (LG) **p < 0.01 compared to agonist treated (HG), n = 4 experiments statistical analysis determined by one-way ANOVA.