Figure 5.

Src phosphorylation in endothelial monolayers treated with p-cresylsulfate (PCS). Human umbilical vein endothelial cell monolayers were treated with 0.1, or 0.2 mM PCS for 2 days. The control group was treated with medium only. Equal amounts (50 µg) of protein samples were loaded in duplicate gels and separated by SDS-PAGE. The protein levels of phosphorylated-Src (pSrc), total Src (Src), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were detected by western blotting. Following electrophoresis, proteins were transferred to polyvinylidene fluoride membranes and detected by either anti-pSrc or anti-Src antibodies via immunostaining on separate membranes. The levels of GAPDH in each membrane were also detected by anti-GAPDH antibodies. (a) Representative results. Lane 1: control sample; lanes 2–3: samples treated with PCS; lane 2: 0.1 mM PCS; and lane 3: 0.2 mM PCS. (b) Protein levels (mean ± SD, n = 3 per group) of pSrc relative to Src.