Figure 3.

Sep-activated wild-type Ras cascade is mediated by NO in HUVEC. A: HUVEC were cultured and pretreated ± Nω-Nitro-L-arginine methyl ester (L-NAME; 10 μmol/L) or LY (10 μmol/L) for one hour. Lysates were then incubated with GST-Raf-RBD. After washing, the pull-down proteins by agarose beads were fractionated by SDS-PAGE and immunoblotted with pan-Ras antibody to GTP-bound Ras and GST-Raf-RBD. B and C: Lysates were also immunoblotted and analyzed for p85α/p110α, p-eNOS (Ser¹¹⁷⁷), p-Akt (Ser⁴⁷³), and GAPDH. D: Schematic diagram of proposing signaling.