Fig. 5. Effects of total saponin from Korean red ginseng (TSKRG) on vasodilator-stimulated phosphoprotein (VASP) phosphorylation of resting or thrombin-stimulated platelets. (A) Effects of TSKRG on phosphorylations of VASP, VASP (Ser¹⁵⁷), VASP (Ser²³⁹). Lane 1, intact platelets (base); lane 2, thrombin; lane 3, thrombin+TSKRG (100 μg/mL); lane 4, thrombin+TSKRG (150 μg/mL). (B) The ratio of VASP (46+50 kDa) to β-actin by TSKRG. (C) The ratio of phosphorylation of VASP (p-VASP) (Ser¹⁵⁷)-50 kDa to β-actin by TSKRG. (D) The ratio of p-VASP (Ser²³⁹)-50 kDa to β-actin by TSKRG. Washed platelets (10⁸/mL) were preincubated with or without TSKRG for 3 min in the presence of 2 mM CaCl₂ and then stimulated with thrombin (0.5 U/mL) for 5 min at 37℃ in an aggregometer. The reactions were terminated by adding an equal volume of lysis buffer. Proteins were extracted, separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) transferred to polyvinylidene difluoride and immunoblotted with the indicated corresponding antidodies, anti-phosphor-VASP Ser²³⁹ or Ser¹⁵⁷. Blots were visualized by ECL plus kit (Amersham, Buckinghamshire, UK) and the immunoblot panels are representative of 3-4 similar experiments. Blots were analyzed by using the Quantity One ver. 4.5 program, and units were expressed density/mm².